| Project/Area Number |
63580206
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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 |
分子遺伝学・分子生理学
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| Research Institution | Osaka University |
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Principal Investigator |
HORII Toshihiro Osaka Univ. Dept.Biology Research associate, 理学部, 助手 (80142305)
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| Co-Investigator(Kenkyū-buntansha) |
OGAWA Tomoko Osaka Univ. Dept.Biology Instructor, 理学部, 講師 (80028208)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1989: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1988: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Homologous DNA recombination / RecA protein / Functional domain / DNA binding / RecA蛋白質 / ドメイン構造 / Dループ形成 / ATPase / SOS機能 |
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| Research Abstract |
The study of in vitro reaction of the RecA protein of Escherichia coli has been playing a frontier role for understanding the molecular mechanism of homologous DNA recombination. In the study funded under the title described above, we focused our efforts on the analyses of molecular structure of the RecA protein to elucidate the molecular mechanism of RecA protein in the strand transfer reaction. In the fiscal years of 1988 and 1989, we obtained the results as described below. 1) Through the analyses of several altered phi8O cI repressor proteins generated by the site directed mutagenesis, it was revealed that the RecA protein does not involved in the catalytic mechanism of the proteolysis of the repressor. The RecA protein enhances the autocleavage activity of the repressor by changing its conformation, suggesting that the RecA protein has any functional donmains for protease. 2) The RecA5327 protein which is deleted 25 amino acids residues from its C-terminal bound to both of single and double stranded DNA with higher affinities than the wild-type RecA protein. Other evidences supporting that the C-terminal region negatively regulates the DNA binding activity of RecA protein were obtained. 3) The region from 40th. to 60th. residues from N-terminal where is rich in hydrophobic amino acid residues functions for RecA-RecA interaction. From these results, we constructed the Head to Head Dimer Model of RecA protein, in which the RecA protein is postulated to contain one DNA binding domain, two domains for RecA-RecA interactions, ATPase domain for expaining the molecular mechanism of RecA protein in forming D-loop.
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