Regulation of functions of RecA protein by the conformational change

通过构象变化调节 RecA 蛋白的功能

• 批准号: 60580212
• 负责人: OGAWA Tomoko
• 金额: $ 1.22万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1985
• 资助国家: 日本
• 起止时间: 1985 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-60580212/
• 关键词: The activated form of RecA protein for the repressor cleavage; RecA protein; <phi> 80 repressor; The cleavage reaction of repressor by RecA protein; a role of the C-terminal region of RecA protein

Purified RecA protein can cleave LexA protein and several prophage repressors ( <lambda> , P22 and 434 ) in vitro at a specific Ala-Gly peptide bonds in the middle of these polypeptide chains. This reaction requires single-stranded DNA, ATP (or ATP- <gamma> -S or dATP) and <Mg^(++)> . To learn what the activated form of RecA protein is and how the cleavage specificity is determined, we investigated the detail mechanisms on the cleavage reaction using <phi> 80, <lambda> and LexA repressors and RecA mutant proteins. Results showed that the RecA-mediated cleavage reaction of the repressors was controled with two processes. One is the formation of an active binary complex between RecA protein and single-stranded DNA. Another is the interaction of the active binary complex with repressor protein. The former process is likely to be controled by the C-terminal region of RecA protein, and the latter process seems to be stimulated by the conformational change of the repressor by interaction with a specific cofactor, such as dGpG or dApG.

纯化的RecA蛋白在体外可<lambda>在这些多肽链中间的一个特异性Ala-Gly肽键上切割莱克萨蛋白和几种前噬菌体阻遏物（P22和434）。该反应需要单链DNA、ATP（或ATP-S<gamma>或dATP）和&lt;Mg^（++）&gt;。为了了解RecA蛋白的激活形式以及切割特异性是如何确定的，我们使用<phi>80<lambda>和莱克萨阻遏物和RecA突变蛋白研究了切割反应的详细机制。结果表明，RecA介导的阻遏物切割反应受两个过程控制。一个是RecA蛋白和单链DNA之间形成活性二元复合物。二是活性二元复合物与阻遏蛋白的相互作用。前一个过程可能受RecA蛋白C-末端区域的控制，后一个过程可能是由阻遏物与特定辅因子（如dGpG或dApG）相互作用而引起的构象变化所刺激的。

项目成果

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小川英行：细胞工程。4. 1197-1208 (1985)

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H. Ogawa: "General recombination: Functions and structure of RecA protein" Advances in Biophysics. 21. 135-148 (1986)

H. Okawa：“一般重组：RecA 蛋白的功能和结构”生物物理学进展。

Y.Eguchi: Journal of Molecular Biology. in Press.

Y.Eguchi：分子生物学杂志。

Y. Eguchi: "Two different forms of RecA protein in cleavage of repressor" Journal of Molecular Biology.

Y. Eguchi：“RecA 蛋白在阻遏物裂解中的两种不同形式”，《分子生物学杂志》。