| Project/Area Number |
03670178
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Experimental pathology
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| Research Institution | Okayama Medical School |
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Principal Investigator |
YOSHIOKA Hidekatsu Okayama Medical School, Molecular biology and Biochemistry, Associate Professor, 医学部, 助教授 (00222430)
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| Co-Investigator(Kenkyū-buntansha) |
NINOMIYA Yoshifumi Okayama Medical School, Molecular biology and Biochemistry, Professor, 医学部, 教授 (70126241)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1993: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1992: ¥200,000 (Direct Cost: ¥200,000)
Fiscal Year 1991: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Collagen / Extracellular Matrix / cDNA / Gene / コラ-ゲン |
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| Research Abstract |
1) Analysis of the promoter of the human alpha1(XI) collagen gene
We have cloned the 5' portion of the human COL11A1 gene and determined the location of its major start site of transcription. Analysis of the immediate upstream sequence revealed the lack of a TATA motif and the presence of several GC boxes. Consistent with this finding, primer extension and RNase protection assays identified several minor transcriptional start sites around a major one, 318-bp upstream from the ATG codon. Transient transfection experiments has showed the 1.6kb upstream region has the ability of the promoter. Deletion constructs narrowed down the sequence of its transcriptional activity between nucleotides -541 to -199. In this region, there is a single foot printing area containing a sequence with similarity to an AP-3 binding site.
2) Primary structure and gene expression of a new human collagen chain
To determine the complete primary structure of the chain, we have isolated overlapping clones from a random-primed cDNA library from human rhabdomyosarcoma cell line (CCL 136) mRNA.The deduced polypeptide contained an open reading frame of 1142 amino acid residues. The predicted polypeptide consists of five collagenous domains of the COL1 to COL5. Structurally, this collagen chain most resembles to the alpha1(XVI) chain. Northern blot analyzes using the 5' end of the cDNA showed 11 mRNA species from 0.7 to 12 kb in size. However, the only two RNAs of 6 and 12kb size out of the multiple RNAs were hybridized to the most 3' end cDNA.The date clearly support the conclusion the cDNAs encode a collagen alpha chain which is different from known alpha chain. We propose to designate this collagen chain as alpha1(XIX) chain.
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