Human COL9A1 and COL9A2 genes. Two genes of 90 and 15 kb code for similar polypeptides of the same collagen molecule

Pihlajamaa, Tero; Vuoristo, Mirka M.; Annunen, Susanna; Perälä, Merja; Prockop, Darwin J.; Ala‐Kokko, Leena

doi:10.1016/s0945-053x(98)90063-4

Cited by 42 publications

(40 citation statements)

References 11 publications

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“…The NC1, NC2, and NC3 domains are of similar size in all three chains, whereas the NC4 domain of the ␣1(IX) chain is much larger than in the ␣2(IX) and ␣3(IX) chains (17, 18). Indeed, the NC4 domain of the ␣1(IX) chain is encoded by exons 1-8 in the COL9A1 gene, which are expressed through the use of the cartilage-specific promoter region studied in the present work (19,20). A second promoter, located between exons 6 and 7, is utilized in other tissues to express a short form of the ␣1(IX) chain that lacks the large NC4 domain (19,20).…”

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confidence: 82%

“…20) and the following primers. For the promoter region: primer DS37, 5Ј-GATCGAATTCAGTAGGG-GGCTTGATGTTA-3Ј (forward) and primer DS38, 5Ј-GATCCTCGAGT-TCCCAGTTGATTTTCTTTG-3Ј (reverse), and for the first intron: primer DS39, 5Ј-GATCGTCGACAAGACAATAACCCTGGAAAGA-3Ј (forward) and primer DS40, 5Ј-GATCAAGCTTTGAAACAGGAGTC-CCCGCAGA-3Ј (reverse).…”

Section: Methodsmentioning

confidence: 99%

“…The primers for the promoter region contained an EcoRI and a XhoI site, respectively, and the primers for the first intron contained a SalI and a HindIII site, respectively, to facilitate cloning. The PCR-generated promoter fragment (976 bp) contained the region from Ϫ846 to ϩ130 relative to the transcriptional start site, and the first intron fragment (807 bp) contained from position ϩ1037 to ϩ1843 in the human COL9A1 gene sequence (excluding the splice-site sequences) (20). The promoter fragment was cloned into the NheI-XhoI sites of the pGL3basic luciferase-reporter vector (Promega, Madison, WI) to generate the full-length 846Luc promoter construct.…”

Section: Methodsmentioning

confidence: 99%

“…Indeed, the NC4 domain of the ␣1(IX) chain is encoded by exons 1-8 in the COL9A1 gene, which are expressed through the use of the cartilage-specific promoter region studied in the present work (19,20). A second promoter, located between exons 6 and 7, is utilized in other tissues to express a short form of the ␣1(IX) chain that lacks the large NC4 domain (19,20). Interestingly, mice that lack type IX collagen develop normally but exhibit a late-onset form of joint degeneration similar to osteoarthritis, suggesting a stabilizing role for type IX collagen in cartilage (21,22).…”

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confidence: 89%

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Regulation of Human COL9A1 Gene Expression

Zhang¹,

Jimenez

Stokes

2003

Journal of Biological Chemistry

111

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The COL9A1 gene contains two promoter regions, one driving expression of a long ␣1(IX) chain in cartilage (upstream) and one driving expression of a shorter chain in the cornea and vitreous (downstream). To determine how the chondrocyte-specific expression of the COL9A1 gene is regulated, we have begun to characterize the upstream chondrocyte-specific promoter region of the human COL9A1 gene. Transient-transfection analyses performed in rat chondrosarcoma (RCS) cells, human chondrosarcoma (HTB) cells, and NIH/3T3 cells showed that the COL9A1 promoter was active in RCS cells but not HTB or NIH/3T3 cells. Inclusion of the first intron had no effect on promoter activity. In transienttransfection analyses with promoter deletion constructs, it was found that full promoter activity in RCS cells depended on the region from ؊560 bp to ؉130 bp relative to the transcriptional start site (؉1). Sequence analysis of the region from ؊890 bp to the transcriptional start predicted five putative SOX/Sry-binding sites. Mutation analysis revealed that two of three putative SOX/Sry binding sites within the ؊560 to ؉130 bp region are responsible for most of the COL9A1 promoter activity in RCS cells. Co-transfection experiments with a SOX9 expression plasmid revealed that a construct containing the five putative SOX/Sry-binding sites was transactivated 20-to 30-fold in both HTB and NIH/3T3 cells. Further co-transfection experiments showed that two of the SOX/Sry-binding sites located within the ؊560 to ؉130 bp region were required for full transactivation. However, mutation and deletion analyses indicated that a region from ؊560 to ؊357 bp, which does not contain any other conspicuous SOX9 sites, is also important for full promoter activity. DNA-protein binding assays and super-shift analysis revealed that SOX9 can form a specific complex with one of the SOX/Sry-binding sites with in the ؊560 to ؉130 region.The chondrocyte is responsible for the precise production of several different types of cartilage tissues in the developing vertebrate; including growth plate cartilage, articular cartilage, and the cartilage of the ear and trachea. In each of these situations, the elaboration of a complex and extensive extracellular matrix, which is the main functional component of cartilaginous tissues, is crucial. The expression of extracellular matrix molecules by chondrocytes must, therefore, be tightly and coordinately controlled at the level of both synthesis and degradation to ensure that the matrix is properly constructed and maintained. Part of this control occurs at the level of the regulation of chondrocyte-specific gene expression. The best studied gene in this regard is the COL2A1 gene, which gives rise to the main fibrillar collagen in the cartilage matrix. The expression of the COL2A1 gene is controlled through transcription factors that interact with both the promoter and the chondrocyte-specific enhancer located within the first intron (1-4). Recent work has shown that both positive and negative factors interact with the COL2A1 g...

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mentioning

confidence: 82%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 89%

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Regulation of Human COL9A1 Gene Expression

Zhang¹,

Jimenez

Stokes

2003

Journal of Biological Chemistry

111

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“…This mobility is consistent with a cross-link between the ␣1(IX)NC1 domain (bearing the 2B4 epitope) and ␣1(II)CB9,7. The COL9A1 sequence encodes a methionine that would be cleaved by CNBr to release a short C-terminal peptide, QAGQRAFNKGPDP, containing the putative cross-linking lysine and the 2B4 epitope (22).…”

Section: Fig 4 Sds-page/mentioning

confidence: 99%

Covalent Cross-linking of the NC1 Domain of Collagen Type IX to Collagen Type II in Cartilage

Eyre

Pietka

Weis

et al. 2004

Journal of Biological Chemistry

120

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From a study to understand the mechanism of covalent interaction between collagen types II and IX, we present experimental evidence for a previously unrecognized molecular site of cross-linking. The location relative to previously defined cross-linking sites predicts a specific manner of interaction and folding of collagen IX on the surface of nascent collagen II fibrils. The initial evidence came from Western blot analysis of type IX collagen extracted by pepsin from fetal human cartilage, which showed a molecular species that had properties indicating an adduct between the ␣1(II) chain and the C-terminal domain (COL1) of type IX collagen. A similar component was isolated from bovine cartilage in sufficient quantity to confirm this identity by N-terminal sequence analysis. Using an antibody that recognized the putative cross-linking sequence at the C terminus of the ␣1(IX) chain, cross-linked peptides were isolated by immunoaffinity chromatography from proteolytic digests of human cartilage collagen. They were characterized by immunochemistry, N-terminal sequence analysis, and mass spectrometry. The results establish a link between a lysine near the C terminus (in the NC1 domain) of ␣1(IX) and the known cross-linking lysine at residue 930 of the ␣1(II) triple helix. This cross-link is speculated to form early in the process of interaction between collagen IX molecules and collagen II polymers. A model of molecular folding and further cross-linking is predicted that can spatially accommodate the formation of all six known cross-linking interactions to the collagen IX molecule on a fibril surface. Of particular biological significance, this model can accommodate potential interfibrillar as well as intrafibrillar links between the collagen IX molecules themselves, so providing a mechanism whereby collagen IX could stabilize a collagen fibril network.Collagen type IX is a member of the fibril-associated collagen with interrupted triple helix (FACIT) 1 family of collagen molecules, which share homologous domains and are all believed to function in the extracellular matrix in association with collagen fibril surfaces (1, 2). Collagen IX is unique among the FACIT molecules in binding covalently to fibril surfaces. The cross-links are formed through the lysyl oxidase mechanism (3-7). Collagen IX is found mostly in cartilages, but it also occurs in the eye (vitreum (8, 9) and avian cornea (8)), ear (tectorial membrane (10)), and intervertebral disc (11, 12), always in co-existence with type II collagen. A special role for collagen IX in the organization of type II collagen fibril networks has apparently evolved, which for articular cartilage seems to be essential for the long term normal functioning of joints. The evidence for this comes from the study of genetic defects. Mutations in all three collagen IX genes have been linked to a chondrodysplasia syndrome (multiple epiphyseal dysplasia), which features early onset osteoarthritis (13).All three chains of the vertebrate collagen IX molecule, ␣1(IX), ␣2(IX), and ␣3(IX)...

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