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: 6933186
• 负责人: Michael M. Cox
• 金额: $ 3.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6933186
• 关键词: Commonwealth of Independent States; DNA damage; DNA repair; Escherichia coli; Pseudomonas aeruginosa; bacterial genetics; bacterial proteins; biochemistry; chimeric proteins; enzyme activity; enzyme inhibitors; enzyme mechanism; enzyme structure; gene deletion mutation; genetic recombination; genetic regulation; hydrolysis; ionizing radiation; microorganism conjugation; mitomycin C; recombinase; ultraviolet radiation

DESCRIPTION (provided by applicant) This research will be done primarily in St. Petersburg, Russia at Petersburg Nuclear Physics Institute in collaboration with Vladislav Lanzov as an extension of NIH grant Number R01GM32335. 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 R01 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 GM32335-21 (recently funded) is to understand how RecA function is autoregulated. Pseudomonas aeruginosa, an important human pathogen, is the source of a RecA protein that we will investigate to further this aim. We propose to continue a productive collaboration that has been extending our understanding of RecA autoregulation through an analysis of the Pseudomonas aeruginosa RecA protein. Work to date has provided numerous mechanistic and structural insights that we will build on. Together with the laboratory of Dr. Vladislava Lanzov in St. Petersburg, Russia, we will further explore the biochemistry of the P. aeruginosa RecA protein, and define the enzymatic differences between it and the E. coli RecA. We have pinpointed likely regions of the protein responsible for autoregulation and for certain other functional differences between the RecA proteins. The new work should help test key features of 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.

描述（由申请人提供）本研究将主要在俄罗斯彼得堡的彼得堡核物理研究所与Vladislav Lanzov合作完成，作为NIH资助号R 01 GM 32335的扩展。重组E.大肠杆菌在体外促进DNA链交换反应，这为重组DNA修复和同源遗传重组的中心步骤提供了方便的分子模型。R 01 GM 32335（考克斯）研究的长期目标是详细了解RecA介导的DNA链交换。重组DNA修复是RecA蛋白在体内的主要功能的假设提供了一个知识框架。GM 32335 -21（最近获得资助）的具体目标之一是了解RecA功能是如何自动调节的。铜绿假单胞菌，一种重要的人类病原体，是RecA蛋白的来源，我们将进一步研究这一目标。我们建议继续进行富有成效的合作，通过对铜绿假单胞菌RecA蛋白的分析，扩展我们对RecA自动调节的理解。迄今为止的工作已经提供了许多机制和结构的见解，我们将建立在与博士Vladislava Lanzov在俄罗斯圣彼得堡彼得堡的实验室，我们将进一步探索铜绿假单胞菌RecA蛋白的生物化学，并确定它和大肠杆菌之间的酶的差异。coli RecA.我们已经确定了负责自动调节和RecA蛋白之间某些其他功能差异的蛋白质的可能区域。这项新工作应该有助于测试RecA介导的DNA链交换模型的关键特征，也可能有助于识别具有增强的DNA结合和链交换功能的RecA变体。这些蛋白质最终可能被证明在基因治疗方案中使用RecA和产生RecA-DNA复合物晶体用于结构分析的努力中是有用的。