2007 Fiscal Year Final Research Report Summary
Analysis of functienal domain and expression in bone, cartilage and muscle of minor collagen
| Project/Area Number |
17390418
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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 |
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Principal Investigator |
YOSHIOKA Hidekatsu Oita University, Faculty of Medicine, Dept. of Anatomy, Biology and Medicine, Professor (00222430)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUO Noritaka Oita University, Faculty of Medicine, Dept. of Anatomy, Biology and Medicine, Assistant Professor (10284788)
SUMIYOSHI Hideaki Oita University, Faculty of Medicine, Dept. of Anatomy, Biology and Medicine, Assistant Professor (60343357)
SHIRABE Komei Oita University, Faculty of Medicine, Dept. of Anatomy, Biology and Medicine, Associate Professor (50179058)
HAMANAKA Ryoji Oita University, Faculty of Medicine, Dept. of Anatomy, Biology and Medicine, Assistant Professor (60274750)
NINOMIYA Yoshifumi Okayama University, Graduate School of Medicine and Dentisuy, Dept. of Molecular Biology and Biochemistry, Professor (70126241)
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| Project Period (FY) |
2005 – 2007
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| Keywords | collagen / transcriptional regulation / bone / cartilage / muscle / extracellular matrix |
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| Research Abstract |
1) Initially, to determine the transcription initiation site of mouse α3 (V) gene, Oligo-Cap methods was used. It was located about 100 bp upstream of ATG codon. We used structure-function analysis of the core promoter region to elucidate the transcriptional features of the gene. The core promoter was defined within about 300 bp immediately upstream from the transcription start site by transient transfection experiments. In this region, we identified two nuclear-factor binding sites by electrophoretic mobility shift assay (EMSAs): BS1 (-130 to -110), BS2 (-190 to -170). Oligonucleotide competition and supershift assays revealed that CBF,/NF-Y specifically bind to BS1.
The N-terminal globular domain of the pro-α3 (V) chain has a unique structure that contains a highly basic segment of 23 amino acids. The peptide showed a specific adhesive feature to osteosarcoma cells but not to chondrosarcoma cells. This feature was derived from four heparin binding sites in the segment. When the peptid
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e adhere to the cell, actin fiber was formed, which was inhibited Rho-kinase inhibitor, Y27632.
2) We have characterized the proximal promoter of the human α1 (III) collagen gene (COL3A1). Transient transfection assays indicated that the segment from -96 to -34 was necessary to activate transcription. EMSAs showed that the multiple proteins form the DNA-protein complex in different combinations depending on the cell types. The region of -79 to -63 bp was critical for DNA protein complex formation. This sequence was contained in the B element of mouse al (III) collagen gene (Col3a1) reported by other researcher.
3) Collagen XXIV is a newly discovered and poorly characterized member of the fibril-forming family of collagen molecules, which is expressed predominantly in bone tissue. Cell transfection experiments in combination with DNA binding assay demonstrated that Co124a1 promoter activity was under control of an upstream cis-acting element, which was recognized by specific combination of c jun, CREB1, ATF1, and ATF2 dimers. Less
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Research Products
(34 results)
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[Journal Article] Elimination of epiplakin by gene targeting results in acceleration of keratinocyte migration in mice2006
Author(s)
Goto, M., Sumiyoshi, H., Sasaki, T., Fassler, R., Ohashi, S., Adachi, E., Yoshioka, H., Fujiwara, S
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Journal Title
Mol. Cell. Biol. 26
Pages: 548-558
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] CREB-AP1 protein complexes regulate transcription of the collagen XXIV gene(Co124a1) in osteoblast2006
Author(s)
Matsuo, N., Tanaka, S., Gordon, M.K., Koch, M., Yoshioka, H., Ramirez, F
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Journal Title
J. Biol. Chem. 281
Pages: 5445-5452
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Multifactor complex containing B element binding factor, BBF, and repressors regulate the human α1(III) collagen gene(COL3A1)2006
Author(s)
Sumiyoshi, H., Matsuo, N., Shin, T., Shirabe, K., Yoshioka, H
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Journal Title
Acta Med. Okayama 60
Pages: 181-189
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Sp1 and Sp3 transcription factors upregulate the proximal promoter of the human prostate-specific antigen(PSA) gene in prostate cancer cells2005
Author(s)
Shin, T., Sumiyoshi, H., Matsuo, N., Satoh, F., Nomura, Y., Mimata, H., Yoshioka, H
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Journal Title
Arch Biochem Biophys 435
Pages: 291-302
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Multiple proteins are involved in the protein-DNA complex in the proximal promoter of the human α1(III) collagen gene(COL3A1)2005
Author(s)
Yoshino, T., Sumiyoshi, H., Shin, T., Matsuo, N., Inagaki, Y., Ninomiya, Y., Yoshioka, H
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Journal Title
Biochimca Biophysica Acta 1729
Pages: 94-104
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Mn-cadherin and its novel variant are transiently expressed in chick embryo spinal cord2005
Author(s)
Shirabe, K., Kimura, Y., Matsuo, N., Fukushima, M., Yoshioka, H., Tanaka, H.
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Journal Title
Biochem Biophys Res Commun 334
Pages: 108-116
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The pro-α3(V) collagen chain is expressed in bone and its N-terminal basic peptide is adhesive to osteosarcoma cells2005
Author(s)
Yamaguchi, K., Matsuo, N., Sumiyoshi, H., Fujimoto, F., Iyama, K., Yanagisawa, S., Yoshioka, H
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Journal Title
Matrix Biol 24
Pages: 283-294
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Adiponectin is expressed in the brown adipose tissue and surrounding immature tissues in mouse embryos2005
Author(s)
Fujimoto, N., Matsuo, N., Sumiyoshi, H., Yamaguchi, K., Saikawa, T., Yoshimatsu, H., Yoshioka, H
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Journal Title
Biochimca Biophysica Acta 1731
Pages: 1-12
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Expression and regulation of pro-α3(V) collagen gene2005
Author(s)
Hidekatsu, Yoshioka, Hitoshi, Nagato, Kenji, Yamaguchi, Ken-ich, Iyama, Keiko, Sakata-Takatani, Hideaki, Sumiyoshi, Noritaka, Matsuo
Organizer
Gordon "Collagen"
Place of Presentation
Colby-Sawyer College, NH, USA
Year and Date
2005-07-25
Description
「研究成果報告書概要(欧文)」より
