Structure/function of RecA protein from P aeruginosa

铜绿假单胞菌 RecA 蛋白的结构/功能

• 批准号: 6540799
• 负责人: Michael M. Cox
• 金额: $ 3.58万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6540799
• 关键词: Commonwealth of Independent States; DNA binding protein; DNA damage; DNA repair; Escherichia coli; Neisseria gonorrhoeae; Pseudomonas aeruginosa; adenosine triphosphate; bacterial genetics; bacterial proteins; biochemistry; chimeric proteins; cooperative study; genetic recombination; hydrolysis; nucleic acid sequence; oligonucleotides; protein structure function

The RecA protein of E. coli promotes a DNA strand exchange reaction in vitro that provides a convenient molecular model for the central steps of recombinational DNA repair and homologous genetic recombination. The long-range goal of the research in ROl GM32335 (Cox) is a detailed understanding of RecA-mediated DNA strand exchange. The hypothesis that recombinational DNA repair is the primary function of RecA protein in vivo provides an intellectual framework. One of the specific aims of GM32335-17 (recently funded) is to carry out a comparative analysis of bacterial RecA proteins and RecA homologues. Pseudomonas aeruginosa, an important human pathogen, is the source of one of the RecA proteins to be investigated. In the present proposal, a collaboration is proposed that should greatly extend our proposed analysis of the Pseudomonas aeruginosa RecA protein, and improve the prospects for mechanistic and structural insights. Together with the laboratory of Dr. Vladislava Lanzov in St. Petersburg, Russia, we will first explore the biochemistry of the P.aeruginosa RecA protein, and establish enzymatic differences between it and the E. coil RecA. Using an extensive set of active chimeric proteins constructed from fusions of the two RecA proteins, we will try to pinpoint regions of the protein responsible for the differences. The work should help test key features of current models for RecA-mediated DNA strand exchange, and may also help identify RecA variants with enhanced DNA binding and strand exchange functions. Such proteins may eventually prove useful in efforts to use RecA in gene therapy protocols and to generate crystals of RecA-DNA complexes for structural analysis.

重组E.大肠杆菌促进体外DNA链交换反应 这为以下核心步骤提供了一个方便的分子模型： 重组DNA修复和同源遗传重组。远程 在ROl GM 32335（考克斯）中的研究的目标是详细了解 RecA介导的DNA链交换。假设重组DNA 修复是RecA蛋白在体内的主要功能， 框架. GM 32335 -17（最近获得资助）的具体目标之一是开展一项 细菌RecA蛋白和RecA同源物的比较分析。 铜绿假单胞菌是一种重要的人类病原体， 要研究的RecA蛋白。在本提案中， 这将大大扩展我们对假单胞菌的分析 铜绿假单胞菌RecA蛋白，并改善机制和前景， 结构性的洞察力。与Vladislava Lanzov博士的实验室一起 俄罗斯的彼得堡，我们将首先探索 铜绿假单胞菌RecA蛋白，并建立其与 急诊线圈RecA。使用一组广泛的活性嵌合蛋白 由两种RecA蛋白的融合构建，我们将尝试精确定位 负责这些差异的蛋白质区域。工作应该会有帮助 测试RecA介导的DNA链交换的当前模型的关键特征，以及 也可以帮助识别具有增强的DNA结合和链的RecA变体 交换功能。这些蛋白质最终可能会被证明是有用的， RecA基因治疗方案和产生RecA-DNA复合物的晶体 进行结构分析。