2004 Fiscal Year Final Research Report Summary
Research on collagen gene expression in chondrocyte and non-chondrocyte, and its function of molecular domains
| Project/Area Number |
14370468
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Oita University (2004)
大分医科大学 (2002-2003) |
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Principal Investigator |
YOSHIOKA Hidekatsu Oita University, Faculty of Medicine, Dept.of Anatomy, Biology and Medicine, Professor, 医学部, 教授 (00222430)
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| Co-Investigator(Kenkyū-buntansha) |
SUMIYOSHI Hideaki Oita University, Faculty of Medicine, Dept.of Anatomy, Biology and Medicine, Assistant Professor, 医学部, 助手 (60343357)
SHIRABE Komei Oita University, Faculty of Medicine, Dept.of Anatomy, Biology and Medicine, Associate Professor, 医学部, 助教授 (50179058)
NINOMIYA Yoshifumi Okayama University, Graduate School of Medicine and Dentistry, Dept.of Molecular Biology and Biochemistry, Professor, 医学部, 教授 (70126241)
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| Project Period (FY) |
2002 – 2004
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| Keywords | collagen / transcriptional regulation / cartilage tissue / bone / extracellular matrix / CBF / NF-Y / V / XI collagen / XIX collagen |
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| Research Abstract |
1)We have characterized the proximal promoter in the human COL11A1, mouse col5α1 and human COL5A3 gene. In COL11A1, a segment from -199 to +1 is necessary for the activation of basal transcriptional regulation. The ATTGG sequence within -147 to -121 is critical for the binding. We identified CBF/NF-Y that bound to the region. In col5α1, the core promoter was defined within the 231 bp upstream from the transcription start site. In this region, we identified three nuclear-factor binding sites. Sp1,CBF/NF-Y, and Sp1-related protein specifically bind to these regions. In these factors, CBF/NF-Y is a key factor in the expression of this gene. In COL5A3, the core promoter was defined within the -129 bp immediately upstream from the transcription start site. In this region, we identified four DNA-protein complexes. The competition assays using consensus oligonucleotides and supershift assays with specific antibodies showed that complex A, which binds to the -122 to -117, also consists of CBF/NF-Y.
2)We examined pro-α3(V) collagen gene expression using immunohistochemical methods combined with in situ hybridization. The pro-α3(V) chain was seen in funis and amnion, but not chorionic villi and deciduas of mouse placenta. In mouse embryo, the transcripts were seen in tissues that were related to bone formation as well as developing muscle and nascent ligament. However, immunohistochemistry showed that pro-α3(V) protein accumulated rather in the developing bone of mouse embryo.
3)We continued characterization of null mice of the collagen XIX gene. Electron microscopy showed thicker BMs around SMC in the gastroesophageal junction.
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