Analysis of the functional domains of a regulatory protein, RecA, using gene manipulation and X-ray crystallography.

使用基因操作和 X 射线晶体学分析调节蛋白 RecA 的功能域。

• 批准号: 59400009
• 负责人: OGAWA Hideyuki
• 金额: $ 11.97万
• 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 (A)
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1985
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-59400009/
• 关键词: RecA蛋白質; 普遍的組換え; 障害修復遺伝子; 蛋白質の構造; recA遺伝子の変異; 機能ドメイン; 抑制変異

For its size (38,000 daltons), the RecA protein exhibits a remarkable variety of reactions in vitro: RecA protein, ( <i> ) binds to a single-stranded DNA; ( <ii> ) has DNAdependent ATPase; ( <iii> ) catalyzes the renaturation of complementary single-strands of DNA; ( <iv> ) unwinds double stranded DNA; and ( <v> ) cleaves LexA protein and the repressors of <lambda> , P22, and <phi> 80 phage. RecA protein probably has several different sites involved directly in these various reactions. To elucidate the molecular mechanism of reactions, location of these reaction sites on the amino acid sequence of the protein would be useful. We therefore first determined the mutation sites of various recA mutants newly isolated by us which are deficient in some or all of the recA functions and, based on the biochemical and biophysical properties of the mutant proteins as well as those of the modified RecA protein, we predicted the functional domains of the RecA protein. To learn how the reactions are done will require determination of the protein's three dimensional structure by X-ray crystallography. We are making effort to obtain a good quality of recA protein crystals suitable for detailed X-ray crystallographic analysis.

就其大小（38,000 道尔顿）而言，RecA 蛋白在体外表现出多种显着的反应：RecA 蛋白 ( <i> ) 与单链 DNA 结合； ( <ii> ) 具有 DNA 依赖性 ATP 酶； ( <iii> ) 催化互补单链 DNA 的复性； ( <iv> ) 解开双链 DNA； ( <v> ) 裂解 LexA 蛋白以及 <lambda>、P22 和 <phi> 80 噬菌体的阻遏蛋白。 RecA 蛋白可能有几个不同的位点直接参与这些不同的反应。为了阐明反应的分子机制，这些反应位点在蛋白质氨基酸序列上的位置将是有用的。因此，我们首先确定了我们新分离的各种recA突变体的突变位点，这些突变体缺乏部分或全部recA功能，并且基于突变体蛋白以及修饰的RecA蛋白的生化和生物物理特性，我们预测了RecA蛋白的功能域。要了解反应如何进行，需要通过 X 射线晶体学测定蛋白质的三维结构。我们正在努力获得适合详细 X 射线晶体学分析的优质 recA 蛋白晶体。

项目成果

Adv. Biophys.21. (1986)

副词。

Biochemistry. 23 - 11. (1984)

生物化学。

Biochemistry. 23-11. (1984)

生物化学。

Proc.Natl.Acad.Sci.USA. 82-13. (1985)

美国国家科学院院刊。

Adv.Biophys.21. (1986)

Adv.Biophys.21。