Functional Organization of DNA Recombinases

DNA 重组酶的功能组织

• 批准号: 6621966
• 负责人: KENDALL L. KNIGHT
• 金额: $ 31.01万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6621966
• 关键词: DNA repair; X ray crystallography; adenosine triphosphate; electron microscopy; enzyme structure; gene mutation; genetic recombination; protein structure function; recombinase; site directed mutagenesis

DESCRIPTION (Provided by applicant): The goal of the work in this proposal is to understand how DNA recombinase enzymes function at the molecular level. Our studies will focus on the bacterial RecA and human Rad5l (HsRad5l) proteins whose activities are of fundamental importance for the maintenance of genomic integrity. In bacteria, RecA is the central regulatory and catalytic component of a DNA-damage inducible recombinational repair system. In humans and other vertebrates, Rad5l is essential for survival and is a central component of a complex set of proteins involved in the catalysis of homologous genetic recombination and recombinational DNA repair. We have recently characterized an unexpected difference between RecA and HsRad5l regarding the regulation of their activity by ATP-mediated allosteric events and will exploit this finding in developing further studies of HsRad5l. We will use a combination of genetic and biochemical methods to address specific mechanistic questions about each enzyme. Studies are also designed to understand how the interaction of HsRad5l with other recombination proteins contributes to its function. Our work addresses questions regarding the molecular nature of conserved aspects of bacterial and human homologous recombination, while at the same time identifying features unique to the human proteins. Understanding the underlying molecular mechanistic principles of such proteins has far-reaching effects in ultimately being able to create novel proteins with desired properties that may be used for prevention of diseases resulting from genomic instability and for beneficial genetic manipulation.

描述（由申请人提供）：本提案中的工作目标是 了解 DNA 重组酶如何在分子水平上发挥作用。我们的 研究将集中于细菌 RecA 和人类 Rad5l (HsRad5l) 蛋白 其活动对于维持基因组至关重要 正直。在细菌中，RecA 是中央调节和催化成分 DNA损伤诱导重组修复系统的研究。在人类和其他 对于脊椎动物来说，Rad5l 对于生存至关重要，并且是生物体的核心组成部分 参与同源遗传催化的一组复杂蛋白质 重组和重组DNA修复。我们最近描述了一个 RecA 和 HsRad5l 在调节方面的意外差异 它们的活性通过 ATP 介导的变构事件进行，并将利用这一发现 开展 HsRad5l 的进一步研究。我们将结合基因 和生化方法来解决每个问题的具体机制问题 酶。研究还旨在了解 HsRad5l 的相互作用如何 与其他重组蛋白的结合有助于其功能。我们的工作 解决了有关保守方面的分子性质的问题 细菌和人类同源重组，同时 识别人类蛋白质独有的特征。了解底层 此类蛋白质的分子机制原理具有深远的影响 最终能够创造出具有所需特性的新型蛋白质 用于预防由基因组不稳定引起的疾病和 有益的基因操纵。