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Gang Bang Gagged. Adult games and cartoons. Preppy amateur girls naked pictures. Lily love reality kings. Viv thomas lesbian workout viv thomas lesbian workout viv thomas lesbian workout viv thomas. MILF Mercedes Carrera Gets Fucked by a BBC. Osteoglycin OGN, a. The SLRP gene family has been conserved since the separation of chondrichthyes and osteichthyes. One exception is ogn for which duplicate copies were identified in fish genomes. The ogn promoter sequence and in vitro mesenchymal stem cell MSC Japan Slrp suggest the duplicate ogn genes acquired divergent functions. In gilthead sea bream Sparus aurata ogn1 was up-regulated during osteoblast and myocyte differentiation in vitro, while ogn2 was severely down-regulated during bone-derived Japan Slrp differentiation into adipocytes in vitro. Japan Slrp, the phylogenetic analysis indicates that the SLRP III family in vertebrates Japan Slrp been under conservative evolutionary pressure. The retention of the ogn gene duplicates https://schalke04fc.info/watersport/video9462-ramuleqy.php teleosts was linked with the acquisition of different functions. Subfunctionalization of Japan Slrp osteoglycin genes ogn1 and ogn2 occurred during teleost evolution. Ogn1 transcripts are Japan Slrp in the early stages of osteoblast and myocyte differentiation in vitro. Ogn2 transcripts are down-regulated in bone-derived MSCs under osteoinductive and adipogenic conditions. The extracellular matrix ECM is important in multicellular organisms and establishes the basic characteristics of each tissue [ 1 ]. The essential building blocks of the ECM are ubiquitous across organisms and include collagens, glycoproteins Japan Slrp proteoglycans [ 2 — 4 ]. The increased ECM complexity in terrestrial and aquatic Japan Slrp relative to early chordates is associated with gene family expansion through duplication of ancestral Japan Slrp genes, and through Japan Slrp small number of vertebrate specific gene innovations [ 1 ]. Knowledge about the ECM in fishes is very patchy despite their unique Japan Slrp and their evolutionary success there are over 34, extant species [ 5 ]. Furthermore, the increased gene number due to teleost specific gene duplications not only elevates the number of potential genes involved in the ECM but also the scope for Japan Slrp innovations [ 6 Japan Slrp 8 ]. They are extracellular proteins with a small protein core, harbouring tandem leucine-rich repeats LRRs that may contain one or more glycosaminoglycan side chains, although there are some exceptions Japan Slrp 910 ]. Nude in front of men Clip download eva maria sex video.

Japanese Mature Milfs. The ogn1 gene from aquatic organisms teleost fish, coelacanth and turtles was under positive selection at several amino acid positions Additional file 7.

Conserved synteny in vertebrate osteoglycins ogn. The gene environment of ogn genes was obtained read more the Ensembl Genome Browser and from the UCSC genome browser of the sea Japan Slrp genome at http: Horizontal lines represent the chromosome fragments and arrow Japan Slrp indicate genes and the arrowhead points in the direction of the predicted gene transcription.

Homologue genes between species are the same colour shading to facilitate perception of conservation. The predicted location of the genes in the chromosome is indicated below each box, in megabase pairs.

Note the higher synteny between OGN in terrestrial vertebrates and teleost ogn1. Japan Slrp searches against the EST https://schalke04fc.info/curly/video11760-fuhi.php in GenBank revealed that transcripts of ogn1 were present in the olfactory epithelium, eye, Japan Slrp, thyroid, skin, bone, scales and digestive tissue while Japan Slrp for ogn2 were detected in jaw, thyroid, thymus, head Japan Slrp, spleen and skeletal muscle of several teleosts C.

Expression profile of gilthead sea bream ogn1 and ogn2 in adult tissues. Quantitative relative expression of a ogn1 and b ogn2 in adult gilthead sea bream tissues. Liver; Gi: Note that muscle, skin and gill arches have the highest relative expression of both ogn1 and 2 in gilthead sea bream. Promoter transcription factors in ogn 1 and ogn 2 genes. Approximately 1. Red rectangles delimit the regions of the promoters that were enriched in chondrocyte, osteoblast, myocyte and adipocyte specific binding sites.

To facilitate identification the different transcription factor binding sites for each cell type Japan Slrp represented in different colours as indicated in the colour chart. Comparison of the transcription factor binding sites in the promoters of sea bass ogn1 and 2 highlighted that the regulation of these genes may explain their different functions in common tissues Fig. Japan Slrp the ogn1 1.

Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSC cultures in osteogenic here. The arrowheads Japan Slrp nodules Japan Slrp mineralization. Normalized expression of b op ; c ogn1 and d ogn2 in bone-derived cells growing Japan Slrp GM Japan Slrp OM at different days of the culture 5 to To determine the osteogenic lineage of cells grown in OM, the expression of the ECM molecule osteopontin op was determined Fig.

Japan Slrp, the expression of ogn1 and ogn2 in cells grown in OM was significantly lower from day 10 to 20 Japan Slrp in Japan Slrp Fig.

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Expression profile of Japan Slrp and ogn2 in gilthead sea bream bone-derived primary MSCs cultures in adipogenic Japan Slrp. Expression profile of ogn1 and ogn2 in gilthead sea bream myocyte primary cultured cells. OGN genes are present in vertebrates, from sharks Japan Slrp mammals and the genome wide gene duplication in teleosts, gave rise to two forms, ogn1 and 2.

The teleost ogn1 and ogn2 gene promoter regions contained common transcription factor binding sites for osteoblasts and myocytes but, while Japan Slrp had binding sites that determine expression in chondrocytes, ogn2 has binding sites that determine expression in adipocytes.

Xxxxwwww Hd Watch Video Chatroulette hot. Note that muscle, skin and gill arches have the highest relative expression of both ogn1 and 2 in gilthead sea bream. Promoter transcription factors in ogn 1 and ogn 2 genes. Approximately 1. Red rectangles delimit the regions of the promoters that were enriched in chondrocyte, osteoblast, myocyte and adipocyte specific binding sites. To facilitate identification the different transcription factor binding sites for each cell type are represented in different colours as indicated in the colour chart. Comparison of the transcription factor binding sites in the promoters of sea bass ogn1 and 2 highlighted that the regulation of these genes may explain their different functions in common tissues Fig. In the ogn1 1. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSC cultures in osteogenic conditions. The arrowheads indicate nodules of mineralization. Normalized expression of b op ; c ogn1 and d ogn2 in bone-derived cells growing in GM or OM at different days of the culture 5 to To determine the osteogenic lineage of cells grown in OM, the expression of the ECM molecule osteopontin op was determined Fig. Conversely, the expression of ogn1 and ogn2 in cells grown in OM was significantly lower from day 10 to 20 than in GM Fig. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSCs cultures in adipogenic conditions. Expression profile of ogn1 and ogn2 in gilthead sea bream myocyte primary cultured cells. OGN genes are present in vertebrates, from sharks to mammals and the genome wide gene duplication in teleosts, gave rise to two forms, ogn1 and 2. The teleost ogn1 and ogn2 gene promoter regions contained common transcription factor binding sites for osteoblasts and myocytes but, while ogn1 had binding sites that determine expression in chondrocytes, ogn2 has binding sites that determine expression in adipocytes. Results of in vitro cultures corroborated the promoter analysis and ogn2 was highly regulated in bone-derived MSC differentiation into adipocytes. The in vitro cell differentiation model and tissue distribution of ogn1 and ogn2 taken together with the gene promoter analysis and divergent motifs in the proteins indicate that subfunctionalization of these duplicated proteoglycans probably occurred in teleosts. In a recent review and classification of SLRPs published in [ 3 ], eighteen SLRP members were identified in the human genome and were clustered into five different classes: Analysis of the SLRP members identified in shark, teleosts, spotted gar, coelacanth and mammals provides further insight into the evolution of proteoglycans in vertebrates. Interestingly, we did not find the NPC gene in the genome of teleost fish, although it was present in other fish genomes including the shark, spotted gar and coelacanth. In the human genome the NPC gene is present on chromosome 6 but is an untranscribed pseudogene, but in other mammals e. Specific gene duplication of ogn was confirmed in all the teleost species analysed. High conservation of the gene environment of ogn1 in teleosts and vertebrates plus the position of teleost ogn1 in the phylogenetic tree suggests it is the orthologue of human OGN. The persistence of duplicate genes in metazoan genomes is quite common [ 67 , 68 ] and it is assumed to be either a consequence of gain of novel function neo-functionalization or partitioning of the function of the ancestral molecule subfunctionalization. Analysis of the phylogenetic tree coupled with the branch-specific test for positive selection indicated that ogn1 is evolving under positive selection in the teleosts, which is coherent with a neo-functionalization model for preservation of gene duplicates [ 69 ]. Overall, evolution has favoured the conservation of this proteoglycan family, despite the major structural e. Thus, it is conceivable that OGN and other SLRP members play crucial functions that are conserved across taxa and that the gene duplicates in teleosts have acquired new functions. Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Conserved clusters of cysteine residues in the N- and C-termini flanked the LRR domains in teleost Ogn1 and Ogn2 and a typical C-terminal leucine-rich repeat cysteine capping motif LRRCE was also present and presumably forms 2 disulphide bridges as observed in mammalian decorin and biglycan [ 64 , 71 , 72 ]. The conservation of the LRRs in teleost Ogns suggest it probably has the curved, solenoid structure revealed by the crystal structure of bovine decorin [ 72 ]. The general conservation of teleost Ogns with mammalian type III proteoglycans suggests that their basic functions are probably conserved. Although the loss of the N-terminal tyrosine sulphate motif in Ogn1 means post-translational addition of keratan sulphate [ 15 ] and the functions resulting from this are unlikely to occur e. To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Interestingly, the first indication of sub-functionalization of ogn duplicates comes from the gilthead sea bream [ 26 , 27 ]. In this species, an increase in Ogn protein and ogn mRNA levels were associated with the regeneration of scales. However, in these damage - repair models, a combination of hard scales and soft tissue damage was also associated with an inflammatory response [ 26 ]. Interestingly, we found 2 promoter modules, containing multiple binding sites for transcription factors that are essential for triggering osteoblast differentiation [ 75 — 78 ] in the ogn1 promoter but not in ogn2 , suggesting the subfunctionalization of these duplicates and a prominent role of ogn1 in this process. In line with this hypothesis, ogn1 mRNA levels increased significantly, as pre-osteoblasts differentiated into osteoblasts in vitro, but ogn2 did not change. Interestingly, when terminal maturation of osteoblasts and active bone mineralization occurred ogn1 and ogn2 mRNA levels were significantly decreased, suggesting that although these ECM related genes might be involved in the development of the scaffold layers of the bone matrix, they do not appear to be essential for its subsequent mineralization. The muscle is a major source of peptides and signalling molecules a. This is the case of OGN that is produced in the skeletal muscle at high levels and have strong bone anabolic effects [ 80 ]. In addition, a role for OGN in muscle differentiation has also been shown using mouse myoblast C2C12 cells undergoing skeletal myogenesis [ 81 ]. In gilthead sea bream, skeletal muscle ogn1 and ogn2 transcripts were highly expressed, although their expression pattern during myocyte differentiation differed. In early stages of myocyte differentiation, the expression of the ogn duplicates does not change. However, ogn1 is up-regulated during myocyte fusion day 8 and formation of myotubes and ogn2 is only modified later day 12 , suggesting it may be important in terminal maturation. Interestingly, these differences may be explained by the different organization in ogn1 and ogn2 promoters. In addition, the involvement of Mrf4 [ 85 ] and OGN [ 86 ] in mammalian muscle cell regeneration has also been described. These coexisting binding sites could reflect the lability of precursor cells that can differentiate into either osteoblasts or myoblasts. Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. These transcription factors are responsible for the induction of the central transcriptional regulators of adipocyte differentiation [ 87 ] and, upon adipocyte differentiation, for the stimulation of transcription of genes involved in lipid biosynthesis and lipid droplet accumulation within the cells [ 88 ]. In this context, we predicted a specific role for ogn2 in adipose tissue. Expression analysis of ogn duplicates in the gilthead seabream bone-derived MSCs undergoing differentiation into adipocyte cells corroborates our predictions as ogn2 expression was significantly down-regulated during this process. The response of ogn2 in these cells agrees with those of mammalian cell lines in which adipocyte cell differentiation from bovine bone marrow cells [ 89 ], mammalian 3T3L1 pre-adipocyte cell line [ 90 ], mouse mesenchymal stem cells MMSCs or senile mouse model-derived bone marrow mesenchymal stem cells SMMSCs [ 91 ] was accompanied by down-regulation of ogn mRNA levels. Thus, in the gilthead seabream, ogn2 is the duplicate that appears to have retained the homologous functions in adipose tissue as described for OGN in mammalians. Characterization of the SLRP members from sharks to mammals indicates that the gene family has been conserved since the separation of chondrichthyes and osteichthyes. Few gene duplications of SLRP members occurred even in the teleosts that suffered a specific whole genome duplication. Analysis of the protein structure of ogn duplicates and the composition of putative transcription binding sites in their gene promoters support the subfunctionalization of these duplicates, which may have favoured the maintenance of duplicate ogn genes in teleosts. Analysis of the ogn promoters together with the in vitro cell culture results indicate that ogn1 is regulated during osteoblast and muscle differentiation up-regulated during osteoblast differentiation and when myocytes start to fuse and form nucleated myotubes, respectively. Conversely, ogn2 appears to be an osteoblast lineage specification factor that is severely down-regulated as cells differentiate into adipocytes and also appears to be involved in the later maturation stages of muscle differentiation when large polynucleated myotubes are formed. Overall our study supports the view that the gene duplicates of ogn in teleosts partitioned functions in the case of myocyte differentiation but also acquired specific functions; ogn1 in osteoblast differentiation and ogn2 as a candidate inhibitor of adipocyte differentiation. Adipogenic medium. Activating transcription factor 4. CCAAT enhancer binding protein alpha. CCAAT enhancer binding protein beta. CCAAT enhancer binding protein delta. Extracellular matrix. Extracellular matrix protein 2. Extracellular matrix protein 2-like. Extracellular matrix protein X. Extracellular matrix protein X-like. Gata binding protein 3. Growth medium. Interferon-stimulated response element. Leucine rich repeats. Myocyte enhancer factor 2. Myocyte enhancer factor 3. Mesenchymal stem cells. Myogenic factor 5. Myogenic factor 6. Myoblast determining factor. Octamer-binding factor 1. Osteogenic medium. Tumor suppressor protein p Podocan and podocan-like. Peroxisome proliferator activated receptor gamma. Proline and arginine rich end leucine rich repeat protein. Runt related transcription factor 2. Runt related transcription factor 3. Small leucine-rich proteoglycans. Sterol regulatory element binding protein. Tsukushi, small leucine rich proteoglycan. The authors would like to thank Emilio J. All authors critically revised the manuscript. All authors read and approved the final manuscript. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Research article Open Access. Vertebrate SLRP family evolution and the subfunctionalization of osteoglycin gene duplicates in teleost fish. Capilla 2 , L. Anjos 1 and D. BMC Evolutionary Biology Results The SLRP gene family has been conserved since the separation of chondrichthyes and osteichthyes. Subfunctionalization of duplicate osteoglycin genes ogn1 and ogn2 occurred during teleost evolution; Ogn1 transcripts are up-regulated in the early stages of osteoblast and myocyte differentiation in vitro; Ogn2 transcripts are down-regulated in bone-derived MSCs under osteoinductive and adipogenic conditions;. Identification and characterization of the osteoglycin ogn gene s in gilthead sea bream To identify homologue s of ogn in the gilthead sea bream S. Phylogenetic analysis and gene environment In this study we first identified the gene repertoire of vertebrate SLRP members in order to, i ensure the correct clustering of gilthead sea bream Ogn sequences, and ii to further characterize when and how ogn genes duplicated in fish. Promoter analysis To assess if the divergent expression of ogn1 and ogn2 was a consequence of divergent regulation at the level of the promoters, the sea bass http: Multiple sequence alignments and protein characterization A multiple sequence alignment ClustalX v2. Myocyte gilthead sea bream primary culture Primary cultures of gilthead sea bream muscle satellite cells were performed as previously described [ 60 ]. A standard curve relating initial template quantity to amplification cycle was generated using serial dilutions of known concentrations of the target template. Transcripts encoding two ogn genes were identified in the gilthead sea bream muscle, vertebra and gill arch transcriptomes Genbank accession numbers: KM and KM for ogn1 and ogn2 , respectively. A multiple sequence alignment of gilthead sea bream Ogn1 and 2 with Ogn from other teleosts, non-teleost fish, amphibians, birds and human revealed a conserved signal peptide sequence and seven characteristic LRR motifs typical of class III SLRP family members Fig. Duplicate genes for Ogn only existed in teleost genomes and presumably arose during the teleost specific whole genome duplication Fig. The teleost Ogns clustered into an Ogn1 and Ogn2 clade and confirmed the identity assigned to the gilthead sea bream ogn1 and 2 cDNAs isolated in this study. The gene-linkage of ogn revealed highly conserved synteny between fish ogn1 and tetrapod OGNs suggesting that it is most like the ancestral form Fig. In contrast, the gene-linkage of fish ogn2 only shared synteny with fish homologues and in zebrafish ogn1 and ogn2 had a single common gene in linkage, namely the duplicated potassium channel tetramerisation domain containing 6 genes kctd6a and kctd6b. Analysis of ogn1 and ogn2 transcript distribution in gilthead sea bream tissues using qPCR corroborated the EST analysis and revealed that ogn1 and ogn2 were highly expressed in muscle, skin and gill arches but were of low abundance in liver, gill filaments, kidney, heart, vertebra and adipose tissue Fig. Head kidney, jaw, thyroid, thymus, spleen, olfactory epithelium, eye and digestive tissue were not analysed by qPCR. UV-responsive transcription factor binding sites have been reported in the human OGN promoter. UV-responsive binding sites were identified in the fish ogn promoters: The morphological analysis of the cells presented in Fig. We further studied the role of ogn duplicates in the process of differentiation of bone-derived MSCs into adipocyte cells by using specific adipogenic conditions AM. Morphological analysis of cells cultured in AM Fig. Representative images of early myoblasts day 2 and small myotubes day 8 are shown in Fig. OGN evolution Specific gene duplication of ogn was confirmed in all the teleost species analysed. OGN structure Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Ogn1 and ogn2 expression in osteoblasts To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Ogn1 and ogn2 expression in myoblast cell cultures The muscle is a major source of peptides and signalling molecules a. Ogn1 and ogn2 expression in bone derived MSCs differentiation into adipocyte cells Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. Adipogenic medium ASPN: Asporin Atf4: Activating transcription factor 4 BGN: Biglycan Cebpa: Contact Temi Oshiyoye, M. Skip to Main Content. You must have Javascript enabled to see this menu. Home About us. JavaScript is not available in your browser. Some enhanced features will not be available until JavaScript is enabled. The SLRP requires a 1: Your basket is currently empty. E3 ubiquitin-protein ligase SlrP. Reviewed - Annotation score: Annotation score: Select a section on the left to see content. This protein is an E3 ubiquitin ligase that interferes with host's ubiquitination pathway. Can ubiquitinate both ubiquitin and host TXN thioredoxin. Leads to significant decrease of thioredoxin activity and increase of host cell death. Cited for: Binding to TXN is inhibited by hydrogen peroxide in vitro. Alternative name s: By similarity. Loss of ubiquitin ligase activity. ModBase i Search MobiDB i Search Several domains are described in this subsection. Belongs to the LRR-containing bacterial E3 ligase family. Mass Da: It is useful for tracking sequence updates. The algorithm is described in the ISO standard. Salmonella enterica subsp. Full view. These are stable identifiers and should be used to cite UniProtKB entries. Q8ZQQ2 Secondary accession number s: March 2, Last sequence update: March 1, Last modified: April 10, This is version 90 of the entry and version 1 of the sequence..

Results Japan Slrp in Japan Slrp cultures Japan Slrp the promoter analysis and ogn2 was highly regulated in bone-derived MSC differentiation into adipocytes.

The in vitro cell differentiation model and tissue distribution of ogn1 and ogn2 Japan Slrp together with the Japan Slrp promoter analysis and divergent motifs in the proteins indicate that subfunctionalization of Japan Slrp duplicated proteoglycans probably occurred in teleosts. In a recent review and classification of SLRPs published in [ 3 ], eighteen SLRP members were identified in the human genome and were clustered into five different classes: Analysis of the SLRP members identified in shark, teleosts, spotted gar, coelacanth and mammals provides further insight into the evolution of proteoglycans in vertebrates.

Interestingly, we did not find the NPC gene in click genome of teleost fish, although it was present in other fish genomes including the shark, spotted gar here coelacanth.

In the human genome the NPC gene is present on chromosome 6 but is an Japan Slrp pseudogene, but in other mammals e. Specific gene duplication of ogn was confirmed in all the teleost species analysed.

Pornbust Xxx Watch Video hot womenxxx. These sequences were then used for phylogenetic analysis using the leucine-rich repeat and immunoglobulin-like domain-containing nogo receptor interacting protein 3 Lingo 3 from Rhincodon typus as an out-group accession number: Phylogenetic analysis was performed using Bayesian inference and the tree was built in MrBayes 3. The accession numbers of all the sequences used to generate the phylogenetic trees are indicated in Additional file 1. A branch-specific test to detect signatures of natural selection in vertebrate OGNs, was used to assess the presence of significantly divergent branches in the ML gene tree Branch Site REL [ 42 ]. The user tree option for analysis in Data Monkey http: To identify the gene environment of fish ogn duplicates and compare it with the gene environment of OGN from other vertebrates, short-range synteny analysis was performed. The genes that flank ogn1 LG1A: To assess if the divergent expression of ogn1 and ogn2 was a consequence of divergent regulation at the level of the promoters, the sea bass http: Position weight matrices were used to represent the transcription factor binding sites using the default parameters. The matrix family library Version 9. A multiple sequence alignment ClustalX v2. The signal peptide, molecular weight and isoelectric point of predicted proteins were determined using SignalP v. Post-translational modification PTM sites were also identified [ 52 — 57 ]. The behaviour and health of all animals was monitored daily and no evidence of infection, modified behaviour or mortality was observed during the experiments. Briefly, the vertebral columns of 6 fish per culture were removed, washed and chopped-up into small fragments with a scalpel. After a week, the bone fragments were removed from the cultures and the adherent cells were collected by treating them with 0. The resulting cell suspension was used to generate several subcultures and cells were maintained for a maximum of 10 passages. For the experiments, cells were trypsinised, suspended in GM, counted and plated in 6-well plates at a density of 10 5 cells per well. Cultures were repeated in 5—8 independent experiments. Primary cultures of gilthead sea bream muscle satellite cells were performed as previously described [ 60 ]. Cells were washed in phosphate buffered saline PBS , resuspended in GM, counted and plated at a density of 1. For cell culture characterisation, images of the cells at different time-points during the experiment were captured with an Axiovert 40C inverted microscope Zeiss coupled to a Canon EOS D digital camera. F forward, R reverse, na not applicable. The longer ogn amplicons were used to generate the standards for qPCR. The results of gene expression analysis were expressed as relative expression copy number for the cell cultures. Significant changes in transcript abundance in the gilthead sea bream tissue panel were tested using a One-way ANOVA with a Bonferroni multiple comparison post-test. Dendrogram comparing OGN structural features from representative organisms of the main vertebrate lineages. The predicted consensus sites for post-translational modifications PTMs are indicated and described in the legend as: Broken lines connect adjacent cysteine pairs and the leucine-rich nuclear export signal is indicated red dashed arrow. The scale above the sequences indicates the amino acid A. A position. The in silico analysis of the molecular weight MW, in kilodaltons, kDa and the isolelectric point Ip of analysed OGNs are indicated on the right hand side of the figure. The accession numbers of the sequences used for structural analysis are given in Additional file 1. Phylogenetic analysis of the vertebrate class III SLRPs revealed 6 main clusters, which contained genes from representatives of the fish species used in the analysis Additional file 5. ECM2 extracellular matrix protein 2 was duplicated in teleosts and the shark C. A further gene cluster that was more like the ancestral gene, ECM2L, was identified in vertebrates but it only contained genes from fish including the shark. The other main branches of the phylogenetic tree contained multiple gene clusters, each of which contained genes from each of the representative species used in the phylogenetic analysis. NPC was absent from teleost fish genomes but present in the shark, spotted gar and coelacanth. Interestingly, although the cysteine motifs were well conserved for each SLRP member across the vertebrates, the motifs were not conserved between SLRP members belonging to the same class e. Phylogenetic relationship of osteoglycins OGNs in vertebrates. Phylogenetic analysis was performed using Bayesian inference and the tree built in MrBayes 3. All other SLRP family members were used to root the tree. The accession number of all the sequences used in this phylogenetic tree are shown in Additional file 1. Ogn from the gar Lepisosteus oculatus was outside the teleost specific Ogn1 and 2 clades. A single OGN homologue was identified in placental mammals, ungulates, rodents, birds, reptiles and amphibians, and in the ancestral fish in the tetrapod lineage, the coelacanth Latimeria chalumnae. The ogn1 gene from aquatic organisms teleost fish, coelacanth and turtles was under positive selection at several amino acid positions Additional file 7. Conserved synteny in vertebrate osteoglycins ogn. The gene environment of ogn genes was obtained from the Ensembl Genome Browser and from the UCSC genome browser of the sea bass genome at http: Horizontal lines represent the chromosome fragments and arrow boxes indicate genes and the arrowhead points in the direction of the predicted gene transcription. Homologue genes between species are the same colour shading to facilitate perception of conservation. The predicted location of the genes in the chromosome is indicated below each box, in megabase pairs. Note the higher synteny between OGN in terrestrial vertebrates and teleost ogn1. BLAST searches against the EST database in GenBank revealed that transcripts of ogn1 were present in the olfactory epithelium, eye, muscle, thyroid, skin, bone, scales and digestive tissue while transcripts for ogn2 were detected in jaw, thyroid, thymus, head kidney, spleen and skeletal muscle of several teleosts C. Expression profile of gilthead sea bream ogn1 and ogn2 in adult tissues. Quantitative relative expression of a ogn1 and b ogn2 in adult gilthead sea bream tissues. Liver; Gi: Note that muscle, skin and gill arches have the highest relative expression of both ogn1 and 2 in gilthead sea bream. Promoter transcription factors in ogn 1 and ogn 2 genes. Approximately 1. Red rectangles delimit the regions of the promoters that were enriched in chondrocyte, osteoblast, myocyte and adipocyte specific binding sites. To facilitate identification the different transcription factor binding sites for each cell type are represented in different colours as indicated in the colour chart. Comparison of the transcription factor binding sites in the promoters of sea bass ogn1 and 2 highlighted that the regulation of these genes may explain their different functions in common tissues Fig. In the ogn1 1. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSC cultures in osteogenic conditions. The arrowheads indicate nodules of mineralization. Normalized expression of b op ; c ogn1 and d ogn2 in bone-derived cells growing in GM or OM at different days of the culture 5 to To determine the osteogenic lineage of cells grown in OM, the expression of the ECM molecule osteopontin op was determined Fig. Conversely, the expression of ogn1 and ogn2 in cells grown in OM was significantly lower from day 10 to 20 than in GM Fig. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSCs cultures in adipogenic conditions. Expression profile of ogn1 and ogn2 in gilthead sea bream myocyte primary cultured cells. OGN genes are present in vertebrates, from sharks to mammals and the genome wide gene duplication in teleosts, gave rise to two forms, ogn1 and 2. The teleost ogn1 and ogn2 gene promoter regions contained common transcription factor binding sites for osteoblasts and myocytes but, while ogn1 had binding sites that determine expression in chondrocytes, ogn2 has binding sites that determine expression in adipocytes. Results of in vitro cultures corroborated the promoter analysis and ogn2 was highly regulated in bone-derived MSC differentiation into adipocytes. The in vitro cell differentiation model and tissue distribution of ogn1 and ogn2 taken together with the gene promoter analysis and divergent motifs in the proteins indicate that subfunctionalization of these duplicated proteoglycans probably occurred in teleosts. In a recent review and classification of SLRPs published in [ 3 ], eighteen SLRP members were identified in the human genome and were clustered into five different classes: Analysis of the SLRP members identified in shark, teleosts, spotted gar, coelacanth and mammals provides further insight into the evolution of proteoglycans in vertebrates. Interestingly, we did not find the NPC gene in the genome of teleost fish, although it was present in other fish genomes including the shark, spotted gar and coelacanth. In the human genome the NPC gene is present on chromosome 6 but is an untranscribed pseudogene, but in other mammals e. Specific gene duplication of ogn was confirmed in all the teleost species analysed. High conservation of the gene environment of ogn1 in teleosts and vertebrates plus the position of teleost ogn1 in the phylogenetic tree suggests it is the orthologue of human OGN. The persistence of duplicate genes in metazoan genomes is quite common [ 67 , 68 ] and it is assumed to be either a consequence of gain of novel function neo-functionalization or partitioning of the function of the ancestral molecule subfunctionalization. Analysis of the phylogenetic tree coupled with the branch-specific test for positive selection indicated that ogn1 is evolving under positive selection in the teleosts, which is coherent with a neo-functionalization model for preservation of gene duplicates [ 69 ]. Overall, evolution has favoured the conservation of this proteoglycan family, despite the major structural e. Thus, it is conceivable that OGN and other SLRP members play crucial functions that are conserved across taxa and that the gene duplicates in teleosts have acquired new functions. Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Conserved clusters of cysteine residues in the N- and C-termini flanked the LRR domains in teleost Ogn1 and Ogn2 and a typical C-terminal leucine-rich repeat cysteine capping motif LRRCE was also present and presumably forms 2 disulphide bridges as observed in mammalian decorin and biglycan [ 64 , 71 , 72 ]. The conservation of the LRRs in teleost Ogns suggest it probably has the curved, solenoid structure revealed by the crystal structure of bovine decorin [ 72 ]. The general conservation of teleost Ogns with mammalian type III proteoglycans suggests that their basic functions are probably conserved. Although the loss of the N-terminal tyrosine sulphate motif in Ogn1 means post-translational addition of keratan sulphate [ 15 ] and the functions resulting from this are unlikely to occur e. To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Interestingly, the first indication of sub-functionalization of ogn duplicates comes from the gilthead sea bream [ 26 , 27 ]. In this species, an increase in Ogn protein and ogn mRNA levels were associated with the regeneration of scales. However, in these damage - repair models, a combination of hard scales and soft tissue damage was also associated with an inflammatory response [ 26 ]. Interestingly, we found 2 promoter modules, containing multiple binding sites for transcription factors that are essential for triggering osteoblast differentiation [ 75 — 78 ] in the ogn1 promoter but not in ogn2 , suggesting the subfunctionalization of these duplicates and a prominent role of ogn1 in this process. In line with this hypothesis, ogn1 mRNA levels increased significantly, as pre-osteoblasts differentiated into osteoblasts in vitro, but ogn2 did not change. Interestingly, when terminal maturation of osteoblasts and active bone mineralization occurred ogn1 and ogn2 mRNA levels were significantly decreased, suggesting that although these ECM related genes might be involved in the development of the scaffold layers of the bone matrix, they do not appear to be essential for its subsequent mineralization. The muscle is a major source of peptides and signalling molecules a. This is the case of OGN that is produced in the skeletal muscle at high levels and have strong bone anabolic effects [ 80 ]. In addition, a role for OGN in muscle differentiation has also been shown using mouse myoblast C2C12 cells undergoing skeletal myogenesis [ 81 ]. In gilthead sea bream, skeletal muscle ogn1 and ogn2 transcripts were highly expressed, although their expression pattern during myocyte differentiation differed. In early stages of myocyte differentiation, the expression of the ogn duplicates does not change. However, ogn1 is up-regulated during myocyte fusion day 8 and formation of myotubes and ogn2 is only modified later day 12 , suggesting it may be important in terminal maturation. Interestingly, these differences may be explained by the different organization in ogn1 and ogn2 promoters. In addition, the involvement of Mrf4 [ 85 ] and OGN [ 86 ] in mammalian muscle cell regeneration has also been described. These coexisting binding sites could reflect the lability of precursor cells that can differentiate into either osteoblasts or myoblasts. Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. These transcription factors are responsible for the induction of the central transcriptional regulators of adipocyte differentiation [ 87 ] and, upon adipocyte differentiation, for the stimulation of transcription of genes involved in lipid biosynthesis and lipid droplet accumulation within the cells [ 88 ]. In this context, we predicted a specific role for ogn2 in adipose tissue. Expression analysis of ogn duplicates in the gilthead seabream bone-derived MSCs undergoing differentiation into adipocyte cells corroborates our predictions as ogn2 expression was significantly down-regulated during this process. The response of ogn2 in these cells agrees with those of mammalian cell lines in which adipocyte cell differentiation from bovine bone marrow cells [ 89 ], mammalian 3T3L1 pre-adipocyte cell line [ 90 ], mouse mesenchymal stem cells MMSCs or senile mouse model-derived bone marrow mesenchymal stem cells SMMSCs [ 91 ] was accompanied by down-regulation of ogn mRNA levels. Thus, in the gilthead seabream, ogn2 is the duplicate that appears to have retained the homologous functions in adipose tissue as described for OGN in mammalians. Characterization of the SLRP members from sharks to mammals indicates that the gene family has been conserved since the separation of chondrichthyes and osteichthyes. Few gene duplications of SLRP members occurred even in the teleosts that suffered a specific whole genome duplication. Analysis of the protein structure of ogn duplicates and the composition of putative transcription binding sites in their gene promoters support the subfunctionalization of these duplicates, which may have favoured the maintenance of duplicate ogn genes in teleosts. Analysis of the ogn promoters together with the in vitro cell culture results indicate that ogn1 is regulated during osteoblast and muscle differentiation up-regulated during osteoblast differentiation and when myocytes start to fuse and form nucleated myotubes, respectively. Conversely, ogn2 appears to be an osteoblast lineage specification factor that is severely down-regulated as cells differentiate into adipocytes and also appears to be involved in the later maturation stages of muscle differentiation when large polynucleated myotubes are formed. Overall our study supports the view that the gene duplicates of ogn in teleosts partitioned functions in the case of myocyte differentiation but also acquired specific functions; ogn1 in osteoblast differentiation and ogn2 as a candidate inhibitor of adipocyte differentiation. Adipogenic medium. Activating transcription factor 4. CCAAT enhancer binding protein alpha. CCAAT enhancer binding protein beta. CCAAT enhancer binding protein delta. Extracellular matrix. Extracellular matrix protein 2. Extracellular matrix protein 2-like. Extracellular matrix protein X. Extracellular matrix protein X-like. Gata binding protein 3. Growth medium. Interferon-stimulated response element. Leucine rich repeats. Myocyte enhancer factor 2. Myocyte enhancer factor 3. Mesenchymal stem cells. ProteinModelPortal i. SMR i. Database of comparative protein structure models More ModBase i. MobiDB i. Linker By similarity. Non-supervised Orthologous Groups More OMA i. Database for complete collections of gene phylogenies More PhylomeDB i. Gene3D i. Integrated resource of protein families, domains and functional sites More InterPro i. Pfam protein domain database More Pfam i. SMART i. These various submissions may originate from different sequencing projects, different types of experiments, or different biological samples. Sequence conflicts are usually of unknown origin. Select the link destinations: EMBL nucleotide sequence database More RefSeq i. Ensembl bacterial and archaeal genome annotation project More EnsemblBacteria i. GeneID i. Kyoto Encyclopedia of Genes and Genomes More KEGG i. ProtoNet; Automatic hierarchical classification of proteins More An eligible practice site could be non-profit or for profit medical facility that 1 treats all persons regardless of their ability to pay and 2 is located in a Health Professional Shortage Area, or 3 Medically Underserved Area, or 4 site is located in one of the 18 rural counties of the State. Physicians and Physician Assistants who are eligible to apply for participation in the SLRP must meet the requirements listed below. Application Deadline and Process: Spring March 1 - April A technical scoring sheet will be used to score and rank applications. The following criteria is scored and ranked when reviewing an application: Award Amount..

High conservation of the gene environment of ogn1 in teleosts and vertebrates plus the position of teleost ogn1 in the phylogenetic tree suggests it is the orthologue of human OGN. The persistence of duplicate genes in metazoan genomes is quite common [ 6768 ] and it is assumed to be either a consequence of Japan Slrp of novel function neo-functionalization or partitioning of the function of the Japan Slrp molecule subfunctionalization. Analysis of the phylogenetic tree coupled with the branch-specific read article for positive selection indicated that ogn1 is evolving under positive selection in the teleosts, which is coherent with a neo-functionalization model for preservation of gene duplicates [ 69 ].

Overall, evolution has favoured the conservation of this proteoglycan family, despite the major structural e. Thus, it is conceivable that OGN and other SLRP members play crucial functions that are conserved across taxa and that the gene duplicates in teleosts have acquired new functions. Japan Slrp Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Conserved clusters of cysteine residues in the Japan Slrp and C-termini flanked the LRR domains Japan Slrp teleost Ogn1 and Ogn2 and a typical C-terminal leucine-rich repeat cysteine capping Japan Slrp LRRCE was also present and presumably forms 2 disulphide bridges as observed in mammalian decorin and biglycan [ 647172 Japan Slrp.

The conservation of Japan Slrp LRRs in teleost Ogns suggest it probably has the curved, solenoid structure revealed by the crystal structure of bovine decorin [ 72 Japan Slrp. The general conservation of teleost Ogns with mammalian type III proteoglycans suggests that their basic functions are probably conserved. Although Japan Slrp loss of the N-terminal tyrosine sulphate motif in Ogn1 means post-translational addition of keratan sulphate [ 15 ] and the functions resulting from this are unlikely to occur e.

To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for Japan Slrp of Ogn function in fish. Interestingly, the first indication of sub-functionalization of ogn duplicates comes from the gilthead sea Japan Slrp [ 2627 Japan Slrp. In this species, Japan Slrp increase in Ogn protein and ogn mRNA levels were Japan Slrp with the regeneration of scales.

However, in these damage - repair models, a combination of hard scales Japan Slrp soft tissue damage was also Japan Slrp with an inflammatory response [ 26 ].

Japan Slrp, we found 2 promoter modules, containing multiple binding sites for transcription factors that are essential for triggering osteoblast differentiation [ 75 — 78 ] in the go here promoter but not in ogn2suggesting the subfunctionalization of these duplicates and a prominent role of Japan Slrp in this process.

In line with this hypothesis, ogn1 mRNA levels increased significantly, as pre-osteoblasts differentiated into osteoblasts in vitro, but ogn2 Japan Slrp not change. Interestingly, when terminal maturation of osteoblasts and Japan Slrp bone mineralization occurred ogn1 and ogn2 mRNA levels were significantly decreased, suggesting that although these ECM related genes might be involved in the Japan Slrp of the scaffold layers of the bone matrix, they do not appear to be essential for its subsequent mineralization.

The muscle is a Poran Sex Video source of peptides and signalling Japan Slrp a. This is the case of OGN that is produced in the skeletal muscle Japan Slrp high levels and have strong bone anabolic effects [ 80 ]. In addition, a role for OGN in muscle differentiation has also been shown using mouse Japan Slrp C2C12 cells undergoing skeletal myogenesis [ 81 ].

In gilthead sea bream, skeletal muscle ogn1 and ogn2 transcripts were highly expressed, although their expression pattern during Japan Slrp differentiation differed. In early stages Japan Slrp myocyte Japan Slrp, the expression of the ogn duplicates does not change. However, ogn1 is up-regulated during myocyte Japan Slrp day 8 and formation of myotubes and ogn2 is only modified later day 12suggesting it may be important in terminal maturation.

Interestingly, these differences may be explained by the Japan Slrp organization in ogn1 and ogn2 promoters. In addition, the involvement of Mrf4 [ 85 ] and OGN [ 86 ] in Japan Slrp muscle cell regeneration has also been described.

Hotwife stories Watch Video Redtube thai. A recipient may reapply for an award after completion of the first 2-year service obligation. The application is currently closed and will reopen on March 1, Contact Temi Oshiyoye, M. Skip to Main Content. You must have Javascript enabled to see this menu. Home About us. JavaScript is not available in your browser. Analysis of ogn1 and ogn2 transcript distribution in gilthead sea bream tissues using qPCR corroborated the EST analysis and revealed that ogn1 and ogn2 were highly expressed in muscle, skin and gill arches but were of low abundance in liver, gill filaments, kidney, heart, vertebra and adipose tissue Fig. Head kidney, jaw, thyroid, thymus, spleen, olfactory epithelium, eye and digestive tissue were not analysed by qPCR. UV-responsive transcription factor binding sites have been reported in the human OGN promoter. UV-responsive binding sites were identified in the fish ogn promoters: The morphological analysis of the cells presented in Fig. We further studied the role of ogn duplicates in the process of differentiation of bone-derived MSCs into adipocyte cells by using specific adipogenic conditions AM. Morphological analysis of cells cultured in AM Fig. Representative images of early myoblasts day 2 and small myotubes day 8 are shown in Fig. OGN evolution Specific gene duplication of ogn was confirmed in all the teleost species analysed. OGN structure Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Ogn1 and ogn2 expression in osteoblasts To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Ogn1 and ogn2 expression in myoblast cell cultures The muscle is a major source of peptides and signalling molecules a. Ogn1 and ogn2 expression in bone derived MSCs differentiation into adipocyte cells Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. Adipogenic medium ASPN: Asporin Atf4: Activating transcription factor 4 BGN: Biglycan Cebpa: Chondroadherin-like DCN: Decorin Dlx1: Extracellular matrix ECM2: Extracellular matrix protein 2 ECM2L: Extracellular matrix protein 2-like ECMX: Epiphycan FMOD: Fibromodulin Gata3: Gata binding protein 3 GM: Growth medium Isre: Interferon-stimulated response element KERA: Keratocan LRR: Leucine rich repeats LUM: Lumican LUML: Lumican-like Mef2: Myocyte enhancer factor 2 Mef3: Myocyte enhancer factor 3 MSCs: Mesenchymal stem cells Msx1—2: Myogenic factor 5 Myf6: Myogenic factor 6 Myod: Myoblast determining factor Nkx3. Nephrocan NYX: Nyctalopin Oct1: Octamer-binding factor 1 OGN: Osteoglycin OM: Osteogenic medium OMD: Osteomodulin Op: Osteopontin OPTC: Opticin p Tumor suppressor protein p53 Pax3: Podocan and podocan-like Pparg: Proline and arginine rich end leucine rich repeat protein Runx2: Runt related transcription factor 2 Runx3: Runt related transcription factor 3 SLRP: Small leucine-rich proteoglycans Sox5: Acknowledgements The authors would like to thank Emilio J. Ethics approval All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. On the origins of the extracellular matrix in vertebrates. Matrix Biol. Functional structure and composition of the extracellular matrix. J Pathol. Proteoglycan form and function: A comprehensive nomenclature of proteoglycans. Biological functions of the small leucine-rich proteoglycans: From genetics to signal transduction. J Biol Chem. Pauly Editors. World Wide Web electronic publication. Whole-genome duplication in teleost fishes and its evolutionary consequences. Mol Genet Genomics. Evolution and diversity of fish genomes. Curr Opin Genet Dev. Comparative analysis of a teleost skeleton transcriptome provides insight into its regulation. Gen Comp Endocrinol. The family of the small leucine-rich proteoglycans: Key regulators of matrix assembly and cellular growth. Crit Rev Biochem Mol. Proteoglycans of the extracellular environment: Clues from the gene and protein side offer novel perspectives in molecular diversity and function. Faseb J. Expression pattern and gene characterization of asporin. The biology of the small leucine-rich proteoglycans - Functional network of interactive proteins. Structural correlations in the family of small leucine-rich repeat proteins and proteoglycans. J Struct Biol. Biglycan knockout mice: New models for musculoskeletal diseases. Glycoconj J. Molecular cloning and tissue distribution of keratocan. Bovine corneal keratan sulfate proteoglycan 37A. Mol Vis. Proteoglycan distribution during healing of corneal stromal wounds in chick. Exp Eye Res. Osteoglycin expression and localization in rabbit tissues and atherosclerotic plaques. Mol Cell Biochem. Identification of osteoglycin as a component of the vascular matrix - Differential expression by vascular smooth muscle cells during neointima formation and in atherosclerotic plaques. Arterioscl Throm Vas. Neurosci Lett. Glucocorticoid up-regulates mimecan expression in corticotroph cells. Mol Cell Endocrinol. Differential expression of mimecan and thioredoxin domain-containing protein 5 in colorectal adenoma and cancer: A proteomic study. Exp Biol Med. Skin and scale regeneration after mechanical damage in a teleost. Mol Immunol. Modulation by Oestradiol beta. Mar Biotechnol. Gene expression in Atlantic salmon skin in response to infection with the parasitic copepod Lepeophtheirus salmonis , cortisol implant, and their combination. BMC Genet. Comparative proteomics analysis of teleost intermuscular bones and ribs provides insight into their development. PLOS One. Development temperature has persistent effects on muscle growth responses in Gilthead sea bream. BMC Bioinf. The Ensembl gene annotation system. European sea bass genome and its variation provide insights into adaptation to euryhalinity and speciation. Nat Commun. Clustal W and clustal X version 2. Nat Genet. New algorithms and methods to estimate maximum-likelihood phylogenies: Syst Biol. MrBayes 3. The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci. Mol Biol Evol. Nucleic Acids Res. MatInspector and beyond: The InterPro protein families database: The Pfam protein families database: SignalP 4. Nat Methods. Walker JM, editor. University of Hertfordshire; Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. EMBO J. Prediction of glycosylation across the human proteome and the correlation to protein function. Analysis and prediction of leucine-rich nuclear export signals. Protein Eng Des Sel. Do not show this banner again. Glycyl thioester intermediate Curated. Ubl conjugation pathway , Virulence. BioCyc i. Recommended name: E3 ubiquitin-protein ligase SlrP EC: Four distinct tokens exist: It lists the nodes as they appear top-down in the taxonomic tree, with the more general grouping listed first. PaxDb, a database of protein abundance averages across all three domains of life More PaxDb i. PRIDE i. Protein interaction database and analysis system More IntAct i. ProteinModelPortal i. SMR i. Database of comparative protein structure models More ModBase i. MobiDB i. Linker By similarity. Non-supervised Orthologous Groups More OMA i. Database for complete collections of gene phylogenies More PhylomeDB i. Gene3D i. Integrated resource of protein families, domains and functional sites More InterPro i. Pfam protein domain database More.

These Japan Slrp binding sites could reflect the lability of precursor cells that can differentiate into either osteoblasts or myoblasts. Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 Very hot anal, the myocyte related transcription factor binding sites coexist with Japan Slrp module of adipocyte related binding sites, which are Japan Slrp from ogn1.

These transcription factors are responsible for the induction of Japan Slrp central transcriptional regulators Japan Slrp adipocyte differentiation [ 87 ] and, upon adipocyte Japan Slrp, for the stimulation of transcription of genes involved in lipid biosynthesis and lipid droplet accumulation within the cells [ 88 ]. In this context, we predicted a specific role for ogn2 in adipose tissue.

Expression analysis of ogn duplicates in the gilthead seabream bone-derived MSCs undergoing differentiation into adipocyte cells corroborates our predictions as ogn2 expression was significantly down-regulated during this process. The response of ogn2 in these cells agrees with those of mammalian cell lines in which adipocyte cell differentiation from bovine bone marrow cells [ 89 Japan Slrp, mammalian 3T3L1 pre-adipocyte cell line [ 90 ], mouse mesenchymal stem cells MMSCs or senile mouse model-derived bone marrow mesenchymal stem cells SMMSCs [ 91 ] was accompanied by down-regulation of ogn mRNA levels.

Thus, in the gilthead seabream, ogn2 is Japan Slrp duplicate that Japan Slrp to source retained Japan Slrp homologous functions in adipose tissue as described for OGN in mammalians. Characterization of the SLRP members Japan Slrp sharks to mammals indicates that the gene family has been conserved since the separation of chondrichthyes and osteichthyes.

Few gene duplications Japan Slrp SLRP members occurred even in the teleosts that suffered Japan Slrp specific whole genome duplication. Analysis of the protein structure of ogn duplicates and the composition of putative transcription binding sites in their gene promoters support the subfunctionalization of these duplicates, which may have favoured the maintenance Japan Slrp duplicate ogn genes in teleosts.

Analysis of the Japan Slrp promoters together with the in vitro cell culture results indicate that ogn1 is regulated during osteoblast and muscle differentiation up-regulated during osteoblast differentiation and when myocytes start to fuse and form Japan Slrp myotubes, Japan Slrp. Conversely, ogn2 appears to be an osteoblast lineage specification factor that is severely down-regulated as cells differentiate into adipocytes and also appears to be involved in the later maturation stages of muscle differentiation when large polynucleated myotubes are formed.

Overall our study supports the view that the gene duplicates of ogn in teleosts partitioned functions in the case of myocyte differentiation but also acquired specific functions; ogn1 Japan Slrp osteoblast differentiation and ogn2 as a candidate inhibitor Japan Slrp adipocyte differentiation. Adipogenic medium.

Sex mri Watch Video Soa Xxxxxhd. Results of in vitro cultures corroborated the promoter analysis and ogn2 was highly regulated in bone-derived MSC differentiation into adipocytes. The in vitro cell differentiation model and tissue distribution of ogn1 and ogn2 taken together with the gene promoter analysis and divergent motifs in the proteins indicate that subfunctionalization of these duplicated proteoglycans probably occurred in teleosts. In a recent review and classification of SLRPs published in [ 3 ], eighteen SLRP members were identified in the human genome and were clustered into five different classes: Analysis of the SLRP members identified in shark, teleosts, spotted gar, coelacanth and mammals provides further insight into the evolution of proteoglycans in vertebrates. Interestingly, we did not find the NPC gene in the genome of teleost fish, although it was present in other fish genomes including the shark, spotted gar and coelacanth. In the human genome the NPC gene is present on chromosome 6 but is an untranscribed pseudogene, but in other mammals e. Specific gene duplication of ogn was confirmed in all the teleost species analysed. High conservation of the gene environment of ogn1 in teleosts and vertebrates plus the position of teleost ogn1 in the phylogenetic tree suggests it is the orthologue of human OGN. The persistence of duplicate genes in metazoan genomes is quite common [ 67 , 68 ] and it is assumed to be either a consequence of gain of novel function neo-functionalization or partitioning of the function of the ancestral molecule subfunctionalization. Analysis of the phylogenetic tree coupled with the branch-specific test for positive selection indicated that ogn1 is evolving under positive selection in the teleosts, which is coherent with a neo-functionalization model for preservation of gene duplicates [ 69 ]. Overall, evolution has favoured the conservation of this proteoglycan family, despite the major structural e. Thus, it is conceivable that OGN and other SLRP members play crucial functions that are conserved across taxa and that the gene duplicates in teleosts have acquired new functions. Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Conserved clusters of cysteine residues in the N- and C-termini flanked the LRR domains in teleost Ogn1 and Ogn2 and a typical C-terminal leucine-rich repeat cysteine capping motif LRRCE was also present and presumably forms 2 disulphide bridges as observed in mammalian decorin and biglycan [ 64 , 71 , 72 ]. The conservation of the LRRs in teleost Ogns suggest it probably has the curved, solenoid structure revealed by the crystal structure of bovine decorin [ 72 ]. The general conservation of teleost Ogns with mammalian type III proteoglycans suggests that their basic functions are probably conserved. Although the loss of the N-terminal tyrosine sulphate motif in Ogn1 means post-translational addition of keratan sulphate [ 15 ] and the functions resulting from this are unlikely to occur e. To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Interestingly, the first indication of sub-functionalization of ogn duplicates comes from the gilthead sea bream [ 26 , 27 ]. In this species, an increase in Ogn protein and ogn mRNA levels were associated with the regeneration of scales. However, in these damage - repair models, a combination of hard scales and soft tissue damage was also associated with an inflammatory response [ 26 ]. Interestingly, we found 2 promoter modules, containing multiple binding sites for transcription factors that are essential for triggering osteoblast differentiation [ 75 — 78 ] in the ogn1 promoter but not in ogn2 , suggesting the subfunctionalization of these duplicates and a prominent role of ogn1 in this process. In line with this hypothesis, ogn1 mRNA levels increased significantly, as pre-osteoblasts differentiated into osteoblasts in vitro, but ogn2 did not change. Interestingly, when terminal maturation of osteoblasts and active bone mineralization occurred ogn1 and ogn2 mRNA levels were significantly decreased, suggesting that although these ECM related genes might be involved in the development of the scaffold layers of the bone matrix, they do not appear to be essential for its subsequent mineralization. The muscle is a major source of peptides and signalling molecules a. This is the case of OGN that is produced in the skeletal muscle at high levels and have strong bone anabolic effects [ 80 ]. In addition, a role for OGN in muscle differentiation has also been shown using mouse myoblast C2C12 cells undergoing skeletal myogenesis [ 81 ]. In gilthead sea bream, skeletal muscle ogn1 and ogn2 transcripts were highly expressed, although their expression pattern during myocyte differentiation differed. In early stages of myocyte differentiation, the expression of the ogn duplicates does not change. However, ogn1 is up-regulated during myocyte fusion day 8 and formation of myotubes and ogn2 is only modified later day 12 , suggesting it may be important in terminal maturation. Interestingly, these differences may be explained by the different organization in ogn1 and ogn2 promoters. In addition, the involvement of Mrf4 [ 85 ] and OGN [ 86 ] in mammalian muscle cell regeneration has also been described. These coexisting binding sites could reflect the lability of precursor cells that can differentiate into either osteoblasts or myoblasts. Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. These transcription factors are responsible for the induction of the central transcriptional regulators of adipocyte differentiation [ 87 ] and, upon adipocyte differentiation, for the stimulation of transcription of genes involved in lipid biosynthesis and lipid droplet accumulation within the cells [ 88 ]. In this context, we predicted a specific role for ogn2 in adipose tissue. Expression analysis of ogn duplicates in the gilthead seabream bone-derived MSCs undergoing differentiation into adipocyte cells corroborates our predictions as ogn2 expression was significantly down-regulated during this process. The response of ogn2 in these cells agrees with those of mammalian cell lines in which adipocyte cell differentiation from bovine bone marrow cells [ 89 ], mammalian 3T3L1 pre-adipocyte cell line [ 90 ], mouse mesenchymal stem cells MMSCs or senile mouse model-derived bone marrow mesenchymal stem cells SMMSCs [ 91 ] was accompanied by down-regulation of ogn mRNA levels. Thus, in the gilthead seabream, ogn2 is the duplicate that appears to have retained the homologous functions in adipose tissue as described for OGN in mammalians. Characterization of the SLRP members from sharks to mammals indicates that the gene family has been conserved since the separation of chondrichthyes and osteichthyes. Few gene duplications of SLRP members occurred even in the teleosts that suffered a specific whole genome duplication. Analysis of the protein structure of ogn duplicates and the composition of putative transcription binding sites in their gene promoters support the subfunctionalization of these duplicates, which may have favoured the maintenance of duplicate ogn genes in teleosts. Analysis of the ogn promoters together with the in vitro cell culture results indicate that ogn1 is regulated during osteoblast and muscle differentiation up-regulated during osteoblast differentiation and when myocytes start to fuse and form nucleated myotubes, respectively. Conversely, ogn2 appears to be an osteoblast lineage specification factor that is severely down-regulated as cells differentiate into adipocytes and also appears to be involved in the later maturation stages of muscle differentiation when large polynucleated myotubes are formed. Overall our study supports the view that the gene duplicates of ogn in teleosts partitioned functions in the case of myocyte differentiation but also acquired specific functions; ogn1 in osteoblast differentiation and ogn2 as a candidate inhibitor of adipocyte differentiation. Adipogenic medium. Activating transcription factor 4. CCAAT enhancer binding protein alpha. CCAAT enhancer binding protein beta. CCAAT enhancer binding protein delta. Extracellular matrix. Extracellular matrix protein 2. Extracellular matrix protein 2-like. Extracellular matrix protein X. Extracellular matrix protein X-like. Gata binding protein 3. Growth medium. Interferon-stimulated response element. Leucine rich repeats. Myocyte enhancer factor 2. Myocyte enhancer factor 3. Mesenchymal stem cells. Myogenic factor 5. Myogenic factor 6. Myoblast determining factor. Octamer-binding factor 1. Osteogenic medium. Tumor suppressor protein p Podocan and podocan-like. Peroxisome proliferator activated receptor gamma. Proline and arginine rich end leucine rich repeat protein. Runt related transcription factor 2. Runt related transcription factor 3. Small leucine-rich proteoglycans. Sterol regulatory element binding protein. Tsukushi, small leucine rich proteoglycan. The authors would like to thank Emilio J. All authors critically revised the manuscript. All authors read and approved the final manuscript. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Research article Open Access. Vertebrate SLRP family evolution and the subfunctionalization of osteoglycin gene duplicates in teleost fish. Capilla 2 , L. Anjos 1 and D. BMC Evolutionary Biology Results The SLRP gene family has been conserved since the separation of chondrichthyes and osteichthyes. Subfunctionalization of duplicate osteoglycin genes ogn1 and ogn2 occurred during teleost evolution; Ogn1 transcripts are up-regulated in the early stages of osteoblast and myocyte differentiation in vitro; Ogn2 transcripts are down-regulated in bone-derived MSCs under osteoinductive and adipogenic conditions;. Identification and characterization of the osteoglycin ogn gene s in gilthead sea bream To identify homologue s of ogn in the gilthead sea bream S. Phylogenetic analysis and gene environment In this study we first identified the gene repertoire of vertebrate SLRP members in order to, i ensure the correct clustering of gilthead sea bream Ogn sequences, and ii to further characterize when and how ogn genes duplicated in fish. Promoter analysis To assess if the divergent expression of ogn1 and ogn2 was a consequence of divergent regulation at the level of the promoters, the sea bass http: Multiple sequence alignments and protein characterization A multiple sequence alignment ClustalX v2. Myocyte gilthead sea bream primary culture Primary cultures of gilthead sea bream muscle satellite cells were performed as previously described [ 60 ]. A standard curve relating initial template quantity to amplification cycle was generated using serial dilutions of known concentrations of the target template. Transcripts encoding two ogn genes were identified in the gilthead sea bream muscle, vertebra and gill arch transcriptomes Genbank accession numbers: KM and KM for ogn1 and ogn2 , respectively. A multiple sequence alignment of gilthead sea bream Ogn1 and 2 with Ogn from other teleosts, non-teleost fish, amphibians, birds and human revealed a conserved signal peptide sequence and seven characteristic LRR motifs typical of class III SLRP family members Fig. Duplicate genes for Ogn only existed in teleost genomes and presumably arose during the teleost specific whole genome duplication Fig. The teleost Ogns clustered into an Ogn1 and Ogn2 clade and confirmed the identity assigned to the gilthead sea bream ogn1 and 2 cDNAs isolated in this study. The gene-linkage of ogn revealed highly conserved synteny between fish ogn1 and tetrapod OGNs suggesting that it is most like the ancestral form Fig. In contrast, the gene-linkage of fish ogn2 only shared synteny with fish homologues and in zebrafish ogn1 and ogn2 had a single common gene in linkage, namely the duplicated potassium channel tetramerisation domain containing 6 genes kctd6a and kctd6b. Analysis of ogn1 and ogn2 transcript distribution in gilthead sea bream tissues using qPCR corroborated the EST analysis and revealed that ogn1 and ogn2 were highly expressed in muscle, skin and gill arches but were of low abundance in liver, gill filaments, kidney, heart, vertebra and adipose tissue Fig. Head kidney, jaw, thyroid, thymus, spleen, olfactory epithelium, eye and digestive tissue were not analysed by qPCR. UV-responsive transcription factor binding sites have been reported in the human OGN promoter. UV-responsive binding sites were identified in the fish ogn promoters: The morphological analysis of the cells presented in Fig. We further studied the role of ogn duplicates in the process of differentiation of bone-derived MSCs into adipocyte cells by using specific adipogenic conditions AM. Morphological analysis of cells cultured in AM Fig. Representative images of early myoblasts day 2 and small myotubes day 8 are shown in Fig. OGN evolution Specific gene duplication of ogn was confirmed in all the teleost species analysed. OGN structure Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Ogn1 and ogn2 expression in osteoblasts To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Ogn1 and ogn2 expression in myoblast cell cultures The muscle is a major source of peptides and signalling molecules a. Ogn1 and ogn2 expression in bone derived MSCs differentiation into adipocyte cells Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. Adipogenic medium ASPN: Asporin Atf4: Activating transcription factor 4 BGN: Biglycan Cebpa: Chondroadherin-like DCN: Decorin Dlx1: Extracellular matrix ECM2: Extracellular matrix protein 2 ECM2L: Extracellular matrix protein 2-like ECMX: Epiphycan FMOD: Fibromodulin Gata3: Gata binding protein 3 GM: Growth medium Isre: Interferon-stimulated response element KERA: Keratocan LRR: Leucine rich repeats LUM: Lumican LUML: Lumican-like Mef2: Myocyte enhancer factor 2 Mef3: Myocyte enhancer factor 3 MSCs: Award Amount. Renewing an Award. A recipient may reapply for an award after completion of the first 2-year service obligation. The application is currently closed and will reopen on March 1, Contact Temi Oshiyoye, M. Skip to Main Content. You must have Javascript enabled to see this menu. Database of comparative protein structure models More ModBase i. MobiDB i. Linker By similarity. Non-supervised Orthologous Groups More OMA i. Database for complete collections of gene phylogenies More PhylomeDB i. Gene3D i. Integrated resource of protein families, domains and functional sites More InterPro i. Pfam protein domain database More Pfam i. SMART i. These various submissions may originate from different sequencing projects, different types of experiments, or different biological samples. Sequence conflicts are usually of unknown origin. Select the link destinations: EMBL nucleotide sequence database More RefSeq i. Ensembl bacterial and archaeal genome annotation project More EnsemblBacteria i. GeneID i. Kyoto Encyclopedia of Genes and Genomes More KEGG i. ProtoNet; Automatic hierarchical classification of proteins More ProtoNet i. Q8ZQQ2 Primary citable accession number:.

Activating transcription factor 4. CCAAT enhancer binding protein alpha. CCAAT enhancer binding protein beta. CCAAT enhancer binding Japan Slrp delta.

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Extracellular matrix. Extracellular matrix protein 2. Japan Slrp matrix protein 2-like. Extracellular matrix protein X. Extracellular matrix protein X-like. Gata binding protein 3. Growth medium. Interferon-stimulated response element. Leucine rich repeats. Myocyte enhancer factor 2. Myocyte enhancer factor Japan Slrp. Mesenchymal stem cells.

Myogenic factor 5.

Odia Girlssex Watch Video Afganu Xxx. Growth medium Isre: Interferon-stimulated response element KERA: Keratocan LRR: Leucine rich repeats LUM: Lumican LUML: Lumican-like Mef2: Myocyte enhancer factor 2 Mef3: Myocyte enhancer factor 3 MSCs: Mesenchymal stem cells Msx1—2: Myogenic factor 5 Myf6: Myogenic factor 6 Myod: Myoblast determining factor Nkx3. Nephrocan NYX: Nyctalopin Oct1: Octamer-binding factor 1 OGN: Osteoglycin OM: Osteogenic medium OMD: Osteomodulin Op: Osteopontin OPTC: Opticin p Tumor suppressor protein p53 Pax3: Podocan and podocan-like Pparg: Proline and arginine rich end leucine rich repeat protein Runx2: Runt related transcription factor 2 Runx3: Runt related transcription factor 3 SLRP: Small leucine-rich proteoglycans Sox5: Acknowledgements The authors would like to thank Emilio J. Ethics approval All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. Consent for publication Not applicable. Competing interests The authors declare that they have no competing interests. On the origins of the extracellular matrix in vertebrates. Matrix Biol. Functional structure and composition of the extracellular matrix. J Pathol. Proteoglycan form and function: A comprehensive nomenclature of proteoglycans. Biological functions of the small leucine-rich proteoglycans: From genetics to signal transduction. J Biol Chem. Pauly Editors. World Wide Web electronic publication. Whole-genome duplication in teleost fishes and its evolutionary consequences. Mol Genet Genomics. Evolution and diversity of fish genomes. Curr Opin Genet Dev. Comparative analysis of a teleost skeleton transcriptome provides insight into its regulation. Gen Comp Endocrinol. The family of the small leucine-rich proteoglycans: Key regulators of matrix assembly and cellular growth. Crit Rev Biochem Mol. Proteoglycans of the extracellular environment: Clues from the gene and protein side offer novel perspectives in molecular diversity and function. Faseb J. Expression pattern and gene characterization of asporin. The biology of the small leucine-rich proteoglycans - Functional network of interactive proteins. Structural correlations in the family of small leucine-rich repeat proteins and proteoglycans. J Struct Biol. Biglycan knockout mice: New models for musculoskeletal diseases. Glycoconj J. Molecular cloning and tissue distribution of keratocan. Bovine corneal keratan sulfate proteoglycan 37A. Mol Vis. Proteoglycan distribution during healing of corneal stromal wounds in chick. Exp Eye Res. Osteoglycin expression and localization in rabbit tissues and atherosclerotic plaques. Mol Cell Biochem. Identification of osteoglycin as a component of the vascular matrix - Differential expression by vascular smooth muscle cells during neointima formation and in atherosclerotic plaques. Arterioscl Throm Vas. Neurosci Lett. Glucocorticoid up-regulates mimecan expression in corticotroph cells. Mol Cell Endocrinol. Differential expression of mimecan and thioredoxin domain-containing protein 5 in colorectal adenoma and cancer: A proteomic study. Exp Biol Med. Skin and scale regeneration after mechanical damage in a teleost. Mol Immunol. Modulation by Oestradiol beta. Mar Biotechnol. Gene expression in Atlantic salmon skin in response to infection with the parasitic copepod Lepeophtheirus salmonis , cortisol implant, and their combination. BMC Genet. Comparative proteomics analysis of teleost intermuscular bones and ribs provides insight into their development. PLOS One. Development temperature has persistent effects on muscle growth responses in Gilthead sea bream. BMC Bioinf. The Ensembl gene annotation system. European sea bass genome and its variation provide insights into adaptation to euryhalinity and speciation. Nat Commun. Clustal W and clustal X version 2. Nat Genet. New algorithms and methods to estimate maximum-likelihood phylogenies: Syst Biol. MrBayes 3. The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci. Mol Biol Evol. Nucleic Acids Res. MatInspector and beyond: The InterPro protein families database: The Pfam protein families database: SignalP 4. Nat Methods. Walker JM, editor. University of Hertfordshire; Prediction of post-translational glycosylation and phosphorylation of proteins from the amino acid sequence. EMBO J. Prediction of glycosylation across the human proteome and the correlation to protein function. Analysis and prediction of leucine-rich nuclear export signals. Protein Eng Des Sel. Incorporating support vector machine for identifying protein tyrosine sulfation sites. J Comput Chem. Prediction of Nepsilon-acetylation on internal lysines implemented in Bayesian Discriminant Method. Biochem Biophys Res Commun. Insulin and IGF-I effects on the proliferation of an osteoblast primary culture from sea bream Sparus aurata. IGF-I binding and receptor signal transduction in primary cell culture of muscle cells of gilthead sea bream: Cell Tissue Res. Divergent responsiveness of the dentary and vertebral bone to a selective estrogen-receptor modulator SERM in the teleost Sparus aurata. Mutations in NYX, encoding the leucine-rich proteoglycan nyctalopin, cause X-linked complete congenital stationary night blindness. A regulatory network controls nephrocan expression and midgut patterning. Preservation of duplicate genes by subfunctionalization. Am Zool. Google Scholar Hughes AL. Adaptive evolution after gene duplication. Trends Genet. Probabilistic cross-species inference of orthologous genomic regions created by whole-genome duplication in yeast. Super-motifs and evolution of tandem leucine-rich repeats within the small proteoglycans--biglycan, decorin, lumican, fibromodulin, PRELP, keratocan, osteoadherin, epiphycan, and osteoglycin. Crystal structure of the biglycan dimer and evidence that dimerization is essential for folding and stability of class I small leucine-rich repeat proteoglycans. Crystal structure of the dimeric protein core of decorin, the archetypal small leucine-rich repeat proteoglycan. Mimecan, the kDa corneal keratan sulfate proteoglycan, is a product of the gene producing osteoglycin. A novel kDa leukocyte-derived osteoglycin enhances the activation of toll-like receptor 4 and exacerbates cardiac inflammation during viral myocarditis. Cell Mol Life Sci. Runx2 overexpression enhances osteoblastic differentiation and mineralization in adipose-derived stem cells in vitro and in vivo. Calcified Tissue Int. Targeted disruption of Cbfa1 results in a complete lack of bone formation owing to maturational arrest of osteoblasts. SMR i. Database of comparative protein structure models More ModBase i. MobiDB i. Linker By similarity. Non-supervised Orthologous Groups More OMA i. Database for complete collections of gene phylogenies More PhylomeDB i. Gene3D i. Integrated resource of protein families, domains and functional sites More InterPro i. Pfam protein domain database More Pfam i. SMART i. These various submissions may originate from different sequencing projects, different types of experiments, or different biological samples. Sequence conflicts are usually of unknown origin. Select the link destinations: EMBL nucleotide sequence database More RefSeq i. Ensembl bacterial and archaeal genome annotation project More EnsemblBacteria i. GeneID i. Kyoto Encyclopedia of Genes and Genomes More KEGG i. ProtoNet; Automatic hierarchical classification of proteins More ProtoNet i. 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Myogenic factor 6. Myoblast determining factor. Octamer-binding factor 1. Osteogenic medium. Tumor suppressor Japan Slrp p Podocan and podocan-like. Peroxisome proliferator activated receptor gamma. Proline and arginine rich end leucine rich Japan Slrp protein. Runt related Japan Slrp factor 2.

Runt related transcription factor 3. Small leucine-rich proteoglycans. Sterol regulatory element binding protein. Tsukushi, small leucine rich proteoglycan. The authors would like to thank Emilio J. All authors critically revised the manuscript. All authors read and approved Japan Slrp final manuscript.

All more info performed in studies involving animals were in accordance with the ethical Japan Slrp of the institution or practice at which the studies were conducted.

Springer Nature remains neutral Japan Slrp regard to jurisdictional claims in published maps and institutional affiliations.

Research article Open Access. Vertebrate SLRP family evolution and the subfunctionalization of osteoglycin gene duplicates in teleost fish. Capilla 2L. Anjos 1 and D.

Turkse sex Watch Video Hot uideo. Physicians and Physician Assistants who are eligible to apply for participation in the SLRP must meet the requirements listed below. Application Deadline and Process: Spring March 1 - April A technical scoring sheet will be used to score and rank applications. The following criteria is scored and ranked when reviewing an application: Award Amount. Renewing an Award. The results of gene expression analysis were expressed as relative expression copy number for the cell cultures. Significant changes in transcript abundance in the gilthead sea bream tissue panel were tested using a One-way ANOVA with a Bonferroni multiple comparison post-test. Dendrogram comparing OGN structural features from representative organisms of the main vertebrate lineages. The predicted consensus sites for post-translational modifications PTMs are indicated and described in the legend as: Broken lines connect adjacent cysteine pairs and the leucine-rich nuclear export signal is indicated red dashed arrow. The scale above the sequences indicates the amino acid A. A position. The in silico analysis of the molecular weight MW, in kilodaltons, kDa and the isolelectric point Ip of analysed OGNs are indicated on the right hand side of the figure. The accession numbers of the sequences used for structural analysis are given in Additional file 1. Phylogenetic analysis of the vertebrate class III SLRPs revealed 6 main clusters, which contained genes from representatives of the fish species used in the analysis Additional file 5. ECM2 extracellular matrix protein 2 was duplicated in teleosts and the shark C. A further gene cluster that was more like the ancestral gene, ECM2L, was identified in vertebrates but it only contained genes from fish including the shark. The other main branches of the phylogenetic tree contained multiple gene clusters, each of which contained genes from each of the representative species used in the phylogenetic analysis. NPC was absent from teleost fish genomes but present in the shark, spotted gar and coelacanth. Interestingly, although the cysteine motifs were well conserved for each SLRP member across the vertebrates, the motifs were not conserved between SLRP members belonging to the same class e. Phylogenetic relationship of osteoglycins OGNs in vertebrates. Phylogenetic analysis was performed using Bayesian inference and the tree built in MrBayes 3. All other SLRP family members were used to root the tree. The accession number of all the sequences used in this phylogenetic tree are shown in Additional file 1. Ogn from the gar Lepisosteus oculatus was outside the teleost specific Ogn1 and 2 clades. A single OGN homologue was identified in placental mammals, ungulates, rodents, birds, reptiles and amphibians, and in the ancestral fish in the tetrapod lineage, the coelacanth Latimeria chalumnae. The ogn1 gene from aquatic organisms teleost fish, coelacanth and turtles was under positive selection at several amino acid positions Additional file 7. Conserved synteny in vertebrate osteoglycins ogn. The gene environment of ogn genes was obtained from the Ensembl Genome Browser and from the UCSC genome browser of the sea bass genome at http: Horizontal lines represent the chromosome fragments and arrow boxes indicate genes and the arrowhead points in the direction of the predicted gene transcription. Homologue genes between species are the same colour shading to facilitate perception of conservation. The predicted location of the genes in the chromosome is indicated below each box, in megabase pairs. Note the higher synteny between OGN in terrestrial vertebrates and teleost ogn1. BLAST searches against the EST database in GenBank revealed that transcripts of ogn1 were present in the olfactory epithelium, eye, muscle, thyroid, skin, bone, scales and digestive tissue while transcripts for ogn2 were detected in jaw, thyroid, thymus, head kidney, spleen and skeletal muscle of several teleosts C. Expression profile of gilthead sea bream ogn1 and ogn2 in adult tissues. Quantitative relative expression of a ogn1 and b ogn2 in adult gilthead sea bream tissues. Liver; Gi: Note that muscle, skin and gill arches have the highest relative expression of both ogn1 and 2 in gilthead sea bream. Promoter transcription factors in ogn 1 and ogn 2 genes. Approximately 1. Red rectangles delimit the regions of the promoters that were enriched in chondrocyte, osteoblast, myocyte and adipocyte specific binding sites. To facilitate identification the different transcription factor binding sites for each cell type are represented in different colours as indicated in the colour chart. Comparison of the transcription factor binding sites in the promoters of sea bass ogn1 and 2 highlighted that the regulation of these genes may explain their different functions in common tissues Fig. In the ogn1 1. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSC cultures in osteogenic conditions. The arrowheads indicate nodules of mineralization. Normalized expression of b op ; c ogn1 and d ogn2 in bone-derived cells growing in GM or OM at different days of the culture 5 to To determine the osteogenic lineage of cells grown in OM, the expression of the ECM molecule osteopontin op was determined Fig. Conversely, the expression of ogn1 and ogn2 in cells grown in OM was significantly lower from day 10 to 20 than in GM Fig. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSCs cultures in adipogenic conditions. Expression profile of ogn1 and ogn2 in gilthead sea bream myocyte primary cultured cells. OGN genes are present in vertebrates, from sharks to mammals and the genome wide gene duplication in teleosts, gave rise to two forms, ogn1 and 2. The teleost ogn1 and ogn2 gene promoter regions contained common transcription factor binding sites for osteoblasts and myocytes but, while ogn1 had binding sites that determine expression in chondrocytes, ogn2 has binding sites that determine expression in adipocytes. Results of in vitro cultures corroborated the promoter analysis and ogn2 was highly regulated in bone-derived MSC differentiation into adipocytes. The in vitro cell differentiation model and tissue distribution of ogn1 and ogn2 taken together with the gene promoter analysis and divergent motifs in the proteins indicate that subfunctionalization of these duplicated proteoglycans probably occurred in teleosts. In a recent review and classification of SLRPs published in [ 3 ], eighteen SLRP members were identified in the human genome and were clustered into five different classes: Analysis of the SLRP members identified in shark, teleosts, spotted gar, coelacanth and mammals provides further insight into the evolution of proteoglycans in vertebrates. Interestingly, we did not find the NPC gene in the genome of teleost fish, although it was present in other fish genomes including the shark, spotted gar and coelacanth. In the human genome the NPC gene is present on chromosome 6 but is an untranscribed pseudogene, but in other mammals e. Specific gene duplication of ogn was confirmed in all the teleost species analysed. High conservation of the gene environment of ogn1 in teleosts and vertebrates plus the position of teleost ogn1 in the phylogenetic tree suggests it is the orthologue of human OGN. The persistence of duplicate genes in metazoan genomes is quite common [ 67 , 68 ] and it is assumed to be either a consequence of gain of novel function neo-functionalization or partitioning of the function of the ancestral molecule subfunctionalization. Analysis of the phylogenetic tree coupled with the branch-specific test for positive selection indicated that ogn1 is evolving under positive selection in the teleosts, which is coherent with a neo-functionalization model for preservation of gene duplicates [ 69 ]. Overall, evolution has favoured the conservation of this proteoglycan family, despite the major structural e. Thus, it is conceivable that OGN and other SLRP members play crucial functions that are conserved across taxa and that the gene duplicates in teleosts have acquired new functions. Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Conserved clusters of cysteine residues in the N- and C-termini flanked the LRR domains in teleost Ogn1 and Ogn2 and a typical C-terminal leucine-rich repeat cysteine capping motif LRRCE was also present and presumably forms 2 disulphide bridges as observed in mammalian decorin and biglycan [ 64 , 71 , 72 ]. The conservation of the LRRs in teleost Ogns suggest it probably has the curved, solenoid structure revealed by the crystal structure of bovine decorin [ 72 ]. The general conservation of teleost Ogns with mammalian type III proteoglycans suggests that their basic functions are probably conserved. Although the loss of the N-terminal tyrosine sulphate motif in Ogn1 means post-translational addition of keratan sulphate [ 15 ] and the functions resulting from this are unlikely to occur e. To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Interestingly, the first indication of sub-functionalization of ogn duplicates comes from the gilthead sea bream [ 26 , 27 ]. In this species, an increase in Ogn protein and ogn mRNA levels were associated with the regeneration of scales. However, in these damage - repair models, a combination of hard scales and soft tissue damage was also associated with an inflammatory response [ 26 ]. Interestingly, we found 2 promoter modules, containing multiple binding sites for transcription factors that are essential for triggering osteoblast differentiation [ 75 — 78 ] in the ogn1 promoter but not in ogn2 , suggesting the subfunctionalization of these duplicates and a prominent role of ogn1 in this process. In line with this hypothesis, ogn1 mRNA levels increased significantly, as pre-osteoblasts differentiated into osteoblasts in vitro, but ogn2 did not change. Interestingly, when terminal maturation of osteoblasts and active bone mineralization occurred ogn1 and ogn2 mRNA levels were significantly decreased, suggesting that although these ECM related genes might be involved in the development of the scaffold layers of the bone matrix, they do not appear to be essential for its subsequent mineralization. The muscle is a major source of peptides and signalling molecules a. This is the case of OGN that is produced in the skeletal muscle at high levels and have strong bone anabolic effects [ 80 ]. In addition, a role for OGN in muscle differentiation has also been shown using mouse myoblast C2C12 cells undergoing skeletal myogenesis [ 81 ]. In gilthead sea bream, skeletal muscle ogn1 and ogn2 transcripts were highly expressed, although their expression pattern during myocyte differentiation differed. In early stages of myocyte differentiation, the expression of the ogn duplicates does not change. However, ogn1 is up-regulated during myocyte fusion day 8 and formation of myotubes and ogn2 is only modified later day 12 , suggesting it may be important in terminal maturation. Interestingly, these differences may be explained by the different organization in ogn1 and ogn2 promoters. In addition, the involvement of Mrf4 [ 85 ] and OGN [ 86 ] in mammalian muscle cell regeneration has also been described. These coexisting binding sites could reflect the lability of precursor cells that can differentiate into either osteoblasts or myoblasts. Interestingly, analysis of teleost ogn promoters also revealed that in ogn2 , the myocyte related transcription factor binding sites coexist with a module of adipocyte related binding sites, which are absent from ogn1. These transcription factors are responsible for the induction of the central transcriptional regulators of adipocyte differentiation [ 87 ] and, upon adipocyte differentiation, for the stimulation of transcription of genes involved in lipid biosynthesis and lipid droplet accumulation within the cells [ 88 ]. In this context, we predicted a specific role for ogn2 in adipose tissue. Expression analysis of ogn duplicates in the gilthead seabream bone-derived MSCs undergoing differentiation into adipocyte cells corroborates our predictions as ogn2 expression was significantly down-regulated during this process. The response of ogn2 in these cells agrees with those of mammalian cell lines in which adipocyte cell differentiation from bovine bone marrow cells [ 89 ], mammalian 3T3L1 pre-adipocyte cell line [ 90 ], mouse mesenchymal stem cells MMSCs or senile mouse model-derived bone marrow mesenchymal stem cells SMMSCs [ 91 ] was accompanied by down-regulation of ogn mRNA levels. Thus, in the gilthead seabream, ogn2 is the duplicate that appears to have retained the homologous functions in adipose tissue as described for OGN in mammalians. Characterization of the SLRP members from sharks to mammals indicates that the gene family has been conserved since the separation of chondrichthyes and osteichthyes. Few gene duplications of SLRP members occurred even in the teleosts that suffered a specific whole genome duplication. Analysis of the protein structure of ogn duplicates and the composition of putative transcription binding sites in their gene promoters support the subfunctionalization of these duplicates, which may have favoured the maintenance of duplicate ogn genes in teleosts. Analysis of the ogn promoters together with the in vitro cell culture results indicate that ogn1 is regulated during osteoblast and muscle differentiation up-regulated during osteoblast differentiation and when myocytes start to fuse and form nucleated myotubes, respectively. Conversely, ogn2 appears to be an osteoblast lineage specification factor that is severely down-regulated as cells differentiate into adipocytes and also appears to be involved in the later maturation stages of muscle differentiation when large polynucleated myotubes are formed. Overall our study supports the view that the gene duplicates of ogn in teleosts partitioned functions in the case of myocyte differentiation but also acquired specific functions; ogn1 in osteoblast differentiation and ogn2 as a candidate inhibitor of adipocyte differentiation. Adipogenic medium. Activating transcription factor 4. CCAAT enhancer binding protein alpha. CCAAT enhancer binding protein beta. CCAAT enhancer binding protein delta. Extracellular matrix. Extracellular matrix protein 2. Extracellular matrix protein 2-like. Extracellular matrix protein X. Extracellular matrix protein X-like. Gata binding protein 3. Growth medium. Interferon-stimulated response element. Leucine rich repeats. Myocyte enhancer factor 2. Myocyte enhancer factor 3. Mesenchymal stem cells. Myogenic factor 5. Myogenic factor 6. Myoblast determining factor. Octamer-binding factor 1. Osteogenic medium. Tumor suppressor protein p Podocan and podocan-like. Peroxisome proliferator activated receptor gamma. Proline and arginine rich end leucine rich repeat protein. Runt related transcription factor 2. Runt related transcription factor 3. Small leucine-rich proteoglycans. Sterol regulatory element binding protein. Tsukushi, small leucine rich proteoglycan. The authors would like to thank Emilio J. All authors critically revised the manuscript. All authors read and approved the final manuscript. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Research article Open Access. Vertebrate SLRP family evolution and the subfunctionalization of osteoglycin gene duplicates in teleost fish. Capilla 2 , L. Anjos 1 and D. BMC Evolutionary Biology Glycyl thioester intermediate Curated. Ubl conjugation pathway , Virulence. BioCyc i. Recommended name: E3 ubiquitin-protein ligase SlrP EC: Four distinct tokens exist: It lists the nodes as they appear top-down in the taxonomic tree, with the more general grouping listed first. PaxDb, a database of protein abundance averages across all three domains of life More PaxDb i. PRIDE i. Protein interaction database and analysis system More IntAct i. ProteinModelPortal i. SMR i. Database of comparative protein structure models More ModBase i. MobiDB i. Linker By similarity. Non-supervised Orthologous Groups More OMA i. Database for complete collections of gene phylogenies More PhylomeDB i. Gene3D i. Integrated resource of protein families, domains and functional sites More InterPro i. Pfam protein domain database More Pfam i..

BMC Evolutionary Biology Results The SLRP gene family has been Japan Slrp since the separation of chondrichthyes and osteichthyes. Subfunctionalization of duplicate osteoglycin genes ogn1 and ogn2 occurred during teleost evolution; Ogn1 transcripts are up-regulated in the Japan Slrp stages of osteoblast and myocyte differentiation in vitro; Ogn2 transcripts are down-regulated in bone-derived MSCs under osteoinductive and adipogenic conditions.

Identification and characterization of the osteoglycin Japan Slrp gene s in gilthead sea bream To identify homologue s of ogn in the gilthead sea bream S. Phylogenetic analysis and gene environment In this study we first identified the gene repertoire of Japan Slrp SLRP members in order to, i ensure the correct Japan Slrp of gilthead sea bream Ogn sequences, and Japan Slrp to further Japan Slrp when and how ogn genes duplicated in fish.

Promoter analysis To assess if the divergent expression of here and ogn2 was a consequence of divergent regulation at the level of the promoters, the sea bass http: Multiple sequence Japan Slrp and protein characterization A multiple sequence alignment ClustalX v2.

E3 ubiquitin-protein ligase SlrP

Myocyte gilthead sea bream primary culture Primary cultures Japan Slrp gilthead sea bream muscle satellite cells were performed as previously described [ 60 ]. A standard curve relating initial template quantity Japan Slrp amplification cycle was generated using serial dilutions of known concentrations of the target template.

Transcripts encoding two ogn genes were identified in the gilthead sea bream muscle, Japan Slrp and gill arch transcriptomes Genbank Japan Slrp numbers: KM and KM for ogn1 and ogn2respectively. A multiple sequence alignment of gilthead sea bream Ogn1 and 2 with Ogn from other Japan Slrp, non-teleost fish, amphibians, birds and human revealed a conserved Japan Slrp peptide sequence and seven characteristic LRR motifs typical of class III SLRP family members Fig.

Duplicate genes for Ogn only existed in teleost genomes and presumably arose during the teleost specific more info genome duplication Fig. The teleost Ogns clustered into an Ogn1 and Ogn2 clade and confirmed the identity assigned to the gilthead sea bream ogn1 and 2 cDNAs Japan Slrp in this study.

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The gene-linkage of ogn revealed highly conserved synteny between fish ogn1 and Japan Slrp OGNs suggesting that it is most like the ancestral form Fig. In contrast, the gene-linkage of fish ogn2 only shared synteny with fish homologues Japan Slrp in Japan Slrp ogn1 and ogn2 Japan Slrp a single common gene in linkage, namely the duplicated potassium channel tetramerisation domain containing 6 genes kctd6a and kctd6b.

Analysis of Japan Slrp and ogn2 transcript distribution in gilthead sea bream tissues using read more corroborated the EST analysis and revealed that ogn1 and ogn2 were highly expressed in Japan Slrp, skin and gill arches but were of low abundance in liver, gill Japan Slrp, kidney, heart, vertebra and adipose tissue Fig.

Head kidney, jaw, thyroid, thymus, spleen, Japan Slrp epithelium, eye and digestive tissue were not analysed by qPCR. UV-responsive transcription factor binding sites have been reported in the human OGN promoter. UV-responsive binding sites were identified in the fish ogn promoters: The Japan Slrp analysis of the cells presented in Fig.

We further studied the role of ogn duplicates in the process of differentiation of bone-derived MSCs into adipocyte cells by using specific adipogenic conditions AM. The algorithm is described in the ISO standard. Japan Slrp enterica subsp. Full view. These are stable identifiers and should be used to cite UniProtKB entries. Japan Slrp Secondary accession number s: March 2, Last sequence Japan Slrp March 1, Last modified: April 10, This is version 90 of the entry and version 1 of the sequence.

See complete history. Main funding by: National Institutes of Health. Do not show this banner again. Glycyl thioester intermediate Curated. Ubl conjugation pathwayVirulence.

Japan Slrp

BioCyc i. Recommended name: E3 ubiquitin-protein ligase SlrP EC: Four distinct tokens exist: It lists the nodes as they appear top-down in the Japan Slrp tree, with the more general grouping listed first.

PaxDb, a database of protein abundance averages across all three domains of life More PaxDb i. PRIDE i. Protein Japan Slrp database and analysis system More IntAct i. ProteinModelPortal i. SMR i. Database of comparative protein structure models More Skip to Main Content. You must have Javascript enabled to see this menu.

Home About Japan Slrp. JavaScript is not available in Japan Slrp browser. Some enhanced features will not be available until JavaScript is enabled. The SLRP requires a 1: Those eligible receive an opportunity Japan Slrp practice in a designated Health Professional Shortage Area HPSA while also Japan Slrp funds to help pay Japan Slrp higher education Japan Slrp. Pokemon porn ash and misty xxx. Systems used to automatically annotate proteins Japan Slrp high accuracy:. Select item s and click on "Add to basket" to create your own collection here entries max.

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Sexy meera Watch Video Download sexx. The application is currently closed and will reopen on March 1, Contact Temi Oshiyoye, M. Skip to Main Content. You must have Javascript enabled to see this menu. Home About us. JavaScript is not available in your browser. Some enhanced features will not be available until JavaScript is enabled. The SLRPs have a diversity of functions that depend on tissue context and the specific characteristics of the organism. Functional compensation can occur between SLRPs and an example of this is the up-regulation of decorin when biglycan is lost in humans [ 14 ]. The present study is focused on osteoglycin OGN, a. OGN KO-mice are viable, fertile and grow normally but the skin has a modified tensile strength due to abnormalities in the collagen fibrils, which are on average thicker [ 17 ]. OGN has a role in wound healing in the cornea, in atherosclerotic lesions and modulates myocardial integrity and remodelling [ 17 — 20 ]. In addition, OGN enhances the neurite outgrowth promoted by insulin-like growth factor-2 and IGF binding protein-2 [ 21 ]. The presence of OGN in the mouse and human pituitary gland co-expressed with proopiomelanocortin and its up-regulation by glucocorticoids and adrenocorticotropic hormone reveals a novel function for OGN in the hypothalamic-pituitary-adrenal axis in mammals [ 22 , 23 ]. An emerging role for OGN secreted by adipose tissue is its action as a satiety factor acting at the level of the hypothalamus [ 24 ]. OGN is also implicated in several pathologies and is down-regulated in tissues derived from colorectal adenomas and cancers when compared to normal mucosa [ 25 ]. Although relatively well characterized in mammals, far less is known about the function of OGN in other vertebrate groups, including fish. In the blunt snout bream Megalobrama amblycephala ogn is involved in the growth of intermuscular bone [ 29 ]. Furthermore, in zebrafish Danio rerio Ogn levels increase significantly during caudal fin regeneration and skeletal development suggesting its involvement in bone and skeletal development [ 30 ]. In the present study we aimed to understand the evolution of teleost ogn in the context of SLRP family evolution. So far, a role for OGN in bone seems to be common across vertebrates but in mammals, several other functions have been described. We characterized gene expression in bone and muscle cell differentiation from gilthead sea bream Sparus aurata mesenchymal stem cells MSCs. The gilthead seabream was chosen for the study as: Overall, the results indicate gene conservation during evolution and retention of the duplicate ogn genes that arose during the teleost specific whole genome duplication. Evidence for subfunctionalization of the duplicate teleost ogn s was uncovered and a role as a candidate factor in early differentiation of multiple cell types was demonstrated. To identify homologue s of ogn in the gilthead sea bream S. OGN sequences were aligned using ClustalX v2. The accession numbers of all the sequences used are indicated in Additional file 1. In this study we first identified the gene repertoire of vertebrate SLRP members in order to, i ensure the correct clustering of gilthead sea bream Ogn sequences, and ii to further characterize when and how ogn genes duplicated in fish. To achieve the first objective, we used the sequences of the 18 known SLRPs in the human genome to search for putative orthologues in sharks Callorhynchus milii and Rhincodon typus , spotted gar Lepisosteus oculatus , African coelacanth Latimeria chalumnae and representatives of teleost fish Cypriniformes: Danio rerio and Perciformes: Dicentrarchus labrax. To improve resolution of the relationship between the ECM like genes we also included the SLRP sequences of other tetrapods Xenopus tropicalis, Anolis carolinensis, Gallus gallus, Mus mus and Sarcophilus harrisii and teleost fish from different orders Pleuronectiformes: Paralichthys olivaceus ; Beloniformes: Oryzias latipes ; Cichliformes: Oreochromis niloticus ; Characiformes: Astyanax mexicanus, Pygocentrus nattereri ; Siluriformes: Ictalurus punctatus ; Salmoniformes: Salmo salar and Cypriniformes: Cyprinus carpio, Sinocyclocheilus graham see Additional file 1 for accession numbers. These sequences were then used for phylogenetic analysis using the leucine-rich repeat and immunoglobulin-like domain-containing nogo receptor interacting protein 3 Lingo 3 from Rhincodon typus as an out-group accession number: Phylogenetic analysis was performed using Bayesian inference and the tree was built in MrBayes 3. The accession numbers of all the sequences used to generate the phylogenetic trees are indicated in Additional file 1. A branch-specific test to detect signatures of natural selection in vertebrate OGNs, was used to assess the presence of significantly divergent branches in the ML gene tree Branch Site REL [ 42 ]. The user tree option for analysis in Data Monkey http: To identify the gene environment of fish ogn duplicates and compare it with the gene environment of OGN from other vertebrates, short-range synteny analysis was performed. The genes that flank ogn1 LG1A: To assess if the divergent expression of ogn1 and ogn2 was a consequence of divergent regulation at the level of the promoters, the sea bass http: Position weight matrices were used to represent the transcription factor binding sites using the default parameters. The matrix family library Version 9. A multiple sequence alignment ClustalX v2. The signal peptide, molecular weight and isoelectric point of predicted proteins were determined using SignalP v. Post-translational modification PTM sites were also identified [ 52 — 57 ]. The behaviour and health of all animals was monitored daily and no evidence of infection, modified behaviour or mortality was observed during the experiments. Briefly, the vertebral columns of 6 fish per culture were removed, washed and chopped-up into small fragments with a scalpel. After a week, the bone fragments were removed from the cultures and the adherent cells were collected by treating them with 0. The resulting cell suspension was used to generate several subcultures and cells were maintained for a maximum of 10 passages. For the experiments, cells were trypsinised, suspended in GM, counted and plated in 6-well plates at a density of 10 5 cells per well. Cultures were repeated in 5—8 independent experiments. Primary cultures of gilthead sea bream muscle satellite cells were performed as previously described [ 60 ]. Cells were washed in phosphate buffered saline PBS , resuspended in GM, counted and plated at a density of 1. For cell culture characterisation, images of the cells at different time-points during the experiment were captured with an Axiovert 40C inverted microscope Zeiss coupled to a Canon EOS D digital camera. F forward, R reverse, na not applicable. The longer ogn amplicons were used to generate the standards for qPCR. The results of gene expression analysis were expressed as relative expression copy number for the cell cultures. Significant changes in transcript abundance in the gilthead sea bream tissue panel were tested using a One-way ANOVA with a Bonferroni multiple comparison post-test. Dendrogram comparing OGN structural features from representative organisms of the main vertebrate lineages. The predicted consensus sites for post-translational modifications PTMs are indicated and described in the legend as: Broken lines connect adjacent cysteine pairs and the leucine-rich nuclear export signal is indicated red dashed arrow. The scale above the sequences indicates the amino acid A. A position. The in silico analysis of the molecular weight MW, in kilodaltons, kDa and the isolelectric point Ip of analysed OGNs are indicated on the right hand side of the figure. The accession numbers of the sequences used for structural analysis are given in Additional file 1. Phylogenetic analysis of the vertebrate class III SLRPs revealed 6 main clusters, which contained genes from representatives of the fish species used in the analysis Additional file 5. ECM2 extracellular matrix protein 2 was duplicated in teleosts and the shark C. A further gene cluster that was more like the ancestral gene, ECM2L, was identified in vertebrates but it only contained genes from fish including the shark. The other main branches of the phylogenetic tree contained multiple gene clusters, each of which contained genes from each of the representative species used in the phylogenetic analysis. NPC was absent from teleost fish genomes but present in the shark, spotted gar and coelacanth. Interestingly, although the cysteine motifs were well conserved for each SLRP member across the vertebrates, the motifs were not conserved between SLRP members belonging to the same class e. Phylogenetic relationship of osteoglycins OGNs in vertebrates. Phylogenetic analysis was performed using Bayesian inference and the tree built in MrBayes 3. All other SLRP family members were used to root the tree. The accession number of all the sequences used in this phylogenetic tree are shown in Additional file 1. Ogn from the gar Lepisosteus oculatus was outside the teleost specific Ogn1 and 2 clades. A single OGN homologue was identified in placental mammals, ungulates, rodents, birds, reptiles and amphibians, and in the ancestral fish in the tetrapod lineage, the coelacanth Latimeria chalumnae. The ogn1 gene from aquatic organisms teleost fish, coelacanth and turtles was under positive selection at several amino acid positions Additional file 7. Conserved synteny in vertebrate osteoglycins ogn. The gene environment of ogn genes was obtained from the Ensembl Genome Browser and from the UCSC genome browser of the sea bass genome at http: Horizontal lines represent the chromosome fragments and arrow boxes indicate genes and the arrowhead points in the direction of the predicted gene transcription. Homologue genes between species are the same colour shading to facilitate perception of conservation. The predicted location of the genes in the chromosome is indicated below each box, in megabase pairs. Note the higher synteny between OGN in terrestrial vertebrates and teleost ogn1. BLAST searches against the EST database in GenBank revealed that transcripts of ogn1 were present in the olfactory epithelium, eye, muscle, thyroid, skin, bone, scales and digestive tissue while transcripts for ogn2 were detected in jaw, thyroid, thymus, head kidney, spleen and skeletal muscle of several teleosts C. Expression profile of gilthead sea bream ogn1 and ogn2 in adult tissues. Quantitative relative expression of a ogn1 and b ogn2 in adult gilthead sea bream tissues. Liver; Gi: Note that muscle, skin and gill arches have the highest relative expression of both ogn1 and 2 in gilthead sea bream. Promoter transcription factors in ogn 1 and ogn 2 genes. Approximately 1. Red rectangles delimit the regions of the promoters that were enriched in chondrocyte, osteoblast, myocyte and adipocyte specific binding sites. To facilitate identification the different transcription factor binding sites for each cell type are represented in different colours as indicated in the colour chart. Comparison of the transcription factor binding sites in the promoters of sea bass ogn1 and 2 highlighted that the regulation of these genes may explain their different functions in common tissues Fig. In the ogn1 1. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSC cultures in osteogenic conditions. The arrowheads indicate nodules of mineralization. Normalized expression of b op ; c ogn1 and d ogn2 in bone-derived cells growing in GM or OM at different days of the culture 5 to To determine the osteogenic lineage of cells grown in OM, the expression of the ECM molecule osteopontin op was determined Fig. Conversely, the expression of ogn1 and ogn2 in cells grown in OM was significantly lower from day 10 to 20 than in GM Fig. Expression profile of ogn1 and ogn2 in gilthead sea bream bone-derived primary MSCs cultures in adipogenic conditions. Expression profile of ogn1 and ogn2 in gilthead sea bream myocyte primary cultured cells. OGN genes are present in vertebrates, from sharks to mammals and the genome wide gene duplication in teleosts, gave rise to two forms, ogn1 and 2. The teleost ogn1 and ogn2 gene promoter regions contained common transcription factor binding sites for osteoblasts and myocytes but, while ogn1 had binding sites that determine expression in chondrocytes, ogn2 has binding sites that determine expression in adipocytes. Results of in vitro cultures corroborated the promoter analysis and ogn2 was highly regulated in bone-derived MSC differentiation into adipocytes. The in vitro cell differentiation model and tissue distribution of ogn1 and ogn2 taken together with the gene promoter analysis and divergent motifs in the proteins indicate that subfunctionalization of these duplicated proteoglycans probably occurred in teleosts. In a recent review and classification of SLRPs published in [ 3 ], eighteen SLRP members were identified in the human genome and were clustered into five different classes: Analysis of the SLRP members identified in shark, teleosts, spotted gar, coelacanth and mammals provides further insight into the evolution of proteoglycans in vertebrates. Interestingly, we did not find the NPC gene in the genome of teleost fish, although it was present in other fish genomes including the shark, spotted gar and coelacanth. In the human genome the NPC gene is present on chromosome 6 but is an untranscribed pseudogene, but in other mammals e. Specific gene duplication of ogn was confirmed in all the teleost species analysed. High conservation of the gene environment of ogn1 in teleosts and vertebrates plus the position of teleost ogn1 in the phylogenetic tree suggests it is the orthologue of human OGN. The persistence of duplicate genes in metazoan genomes is quite common [ 67 , 68 ] and it is assumed to be either a consequence of gain of novel function neo-functionalization or partitioning of the function of the ancestral molecule subfunctionalization. Analysis of the phylogenetic tree coupled with the branch-specific test for positive selection indicated that ogn1 is evolving under positive selection in the teleosts, which is coherent with a neo-functionalization model for preservation of gene duplicates [ 69 ]. Overall, evolution has favoured the conservation of this proteoglycan family, despite the major structural e. Thus, it is conceivable that OGN and other SLRP members play crucial functions that are conserved across taxa and that the gene duplicates in teleosts have acquired new functions. Teleost Ogn duplicates possess most of the key structural motifs that characterize the proteoglycans: Conserved clusters of cysteine residues in the N- and C-termini flanked the LRR domains in teleost Ogn1 and Ogn2 and a typical C-terminal leucine-rich repeat cysteine capping motif LRRCE was also present and presumably forms 2 disulphide bridges as observed in mammalian decorin and biglycan [ 64 , 71 , 72 ]. The conservation of the LRRs in teleost Ogns suggest it probably has the curved, solenoid structure revealed by the crystal structure of bovine decorin [ 72 ]. The general conservation of teleost Ogns with mammalian type III proteoglycans suggests that their basic functions are probably conserved. Although the loss of the N-terminal tyrosine sulphate motif in Ogn1 means post-translational addition of keratan sulphate [ 15 ] and the functions resulting from this are unlikely to occur e. To assess if teleost Ogn duplicates underwent subfunctionalization, as suggested by the promoter analysis in the present study, we searched for reports of Ogn function in fish. Interestingly, the first indication of sub-functionalization of ogn duplicates comes from the gilthead sea bream [ 26 , 27 ]. In this species, an increase in Ogn protein and ogn mRNA levels were associated with the regeneration of scales. However, in these damage - repair models, a combination of hard scales and soft tissue damage was also associated with an inflammatory response [ 26 ]. Interestingly, we found 2 promoter modules, containing multiple binding sites for transcription factors that are essential for triggering osteoblast differentiation [ 75 — 78 ] in the ogn1 promoter but not in ogn2 , suggesting the subfunctionalization of these duplicates and a prominent role of ogn1 in this process. In line with this hypothesis, ogn1 mRNA levels increased significantly, as pre-osteoblasts differentiated into osteoblasts in vitro, but ogn2 did not change. Interestingly, when terminal maturation of osteoblasts and active bone mineralization occurred ogn1 and ogn2 mRNA levels were significantly decreased, suggesting that although these ECM related genes might be involved in the development of the scaffold layers of the bone matrix, they do not appear to be essential for its subsequent mineralization. The muscle is a major source of peptides and signalling molecules a. This is the case of OGN that is produced in the skeletal muscle at high levels and have strong bone anabolic effects [ 80 ]. In addition, a role for OGN in muscle differentiation has also been shown using mouse myoblast C2C12 cells undergoing skeletal myogenesis [ 81 ]. In gilthead sea bream, skeletal muscle ogn1 and ogn2 transcripts were highly expressed, although their expression pattern during myocyte differentiation differed. In early stages of myocyte differentiation, the expression of the ogn duplicates does not change. Systems used to automatically annotate proteins with high accuracy:. Select item s and click on "Add to basket" to create your own collection here entries max. Manual assertion based on experiment in i. You are using a version of browser that may not display all the features of this website. Please consider upgrading your browser. Basket 0. Your basket is currently empty. E3 ubiquitin-protein ligase SlrP. Reviewed - Annotation score: Annotation score: Select a section on the left to see content. This protein is an E3 ubiquitin ligase that interferes with host's ubiquitination pathway. Can ubiquitinate both ubiquitin and host TXN thioredoxin. Leads to significant decrease of thioredoxin activity and increase of host cell death. Cited for: Binding to TXN is inhibited by hydrogen peroxide in vitro. Alternative name s: By similarity. Loss of ubiquitin ligase activity. ModBase i Search MobiDB i Search Several domains are described in this subsection. Belongs to the LRR-containing bacterial E3 ligase family. Mass Da: It is useful for tracking sequence updates. The algorithm is described in the ISO standard. Salmonella enterica subsp..

Your basket is currently empty. E3 ubiquitin-protein ligase SlrP. Japan Slrp - Japan Slrp score: Annotation score: Select a section on the left to see content. This protein is an E3 ubiquitin ligase that interferes with host's ubiquitination pathway.

Can ubiquitinate both ubiquitin and host TXN thioredoxin. Leads to significant decrease of thioredoxin activity and Japan Slrp of host cell death. Cited for: Binding to TXN is inhibited by hydrogen peroxide in vitro. Alternative name Japan Slrp By similarity. Loss of ubiquitin ligase activity. ModBase i Search MobiDB i Search Several Japan Slrp are described in this subsection.

Belongs to the LRR-containing bacterial E3 ligase family. Mass Japan Slrp It is useful for tracking sequence updates. The Japan Slrp is described in the ISO standard. Salmonella enterica subsp. Full view. These are stable identifiers and should be used to cite UniProtKB entries. Q8ZQQ2 Secondary accession number s: March 2, Last sequence update: March 1, Last modified: April 10, This is version 90 of the entry and version 1 of the sequence.

See complete history. Main funding by: National Institutes of Health. Do not show this banner again. Japan Slrp thioester intermediate Curated. Ubl conjugation Japan SlrpVirulence. BioCyc i. Recommended name: E3 ubiquitin-protein ligase SlrP EC: Four distinct tokens exist: It lists Japan Slrp nodes as they appear top-down in the taxonomic tree, with the more general grouping listed first.

PaxDb, a database of protein abundance averages learn more here all three domains of life More PaxDb i. PRIDE i. Protein interaction database and analysis system More IntAct i. Japan Slrp i. SMR i. Database of comparative protein structure models More ModBase i. MobiDB i. Linker By similarity. Non-supervised Orthologous Groups More OMA i. Database for complete collections of gene phylogenies More PhylomeDB i.

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Gene3D i. Integrated resource of protein families, domains and functional sites More InterPro i. Japan Slrp protein domain Japan Slrp More Pfam i. SMART i. These various submissions may originate from different sequencing projects, different Japan Slrp of experiments, or different biological samples.

Sequence conflicts are usually of unknown origin. Select the link destinations: EMBL nucleotide sequence database More RefSeq i. Ensembl bacterial and archaeal genome annotation project More EnsemblBacteria i.

GeneID i. Kyoto Encyclopedia of Genes and Genomes More KEGG i. ProtoNet; Automatic Japan Slrp classification of proteins More ProtoNet i. Japan Slrp Primary citable accession number: This is version 90 of the entry and see more 1 of the sequence. Annotation program. Prokaryotic Protein Annotation Program. I wanna butt fuck.

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