Structure and Function of Bacterial RecQ Protein

细菌 RecQ 蛋白的结构和功能

• 批准号: 6888495
• 负责人: James L Keck
• 金额: $ 25.04万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6888495
• 关键词: DNA repair; DNA replication; X ray crystallography; bacterial proteins; chemical stability; clinical research; enzyme mechanism; genetic recombination; helicase; isozymes; protein binding; protein protein interaction; protein quantitation /detection; protein structure function; structural biology

DESCRIPTION (provided by applicant): The goal of this proposal is to determine the molecular mechanisms used by RecQ DNA helicases. RecQ involvement in DNA replication, recombination, and repair has been demonstrated in both bacteria and eukaryotes, but the precise roles played by this family of helicases remain unclear. Their importance, however, is unambiguous -- mutation of one of three human RecQ genes leads to genome instability and an elevated predisposition to cancer in Werner's, Bloom's, and Rothmund-Thompson syndromes as well as to premature aging in Werner's and Rothmund-Thompson syndromes. RecQ proteins, therefore, represent a paradigm linking genome stability to disease states as well as for understanding the connections between DNA replication, recombination, and repair. This makes a precise, molecular understanding of the mechanisms of the RecQ family important to both basic and medical science. The experiments described in this proposal aim to elucidate the biological functions of RecQ proteins by resolving their molecular mechanisms of action in vitro and in viva. The approach focuses biochemical, high-resolution structural, and genetic methods on bacterial RecQ proteins to delineate the structure-function relationship that supports activity in the enzyme. These experiments include: (1) a determination of the roles of the conserved domains within the RecQ family (helicase, RecQ-Ct, and HRDC domains), (2) elucidation of the quaternary and tertiary structures of RecQ, and (3) identification of RecQ-containing protein complexes in bacterial cells and a study of the effects of interacting proteins on RecQ activity in vitro. A multidisciplinary approach will allow biochemical links to be made between the cellular roles of RecQ proteins and their molecular mechanisms of action.

描述（由申请人提供）： 该建议的目的是确定RECQ DNA解旋酶使用的分子机制。在细菌和真核生物中都证明了RECQ参与DNA复制，重组和修复，但是这种解旋酶家族的确切作用尚不清楚。然而，它们的重要性是明确的 - 三种人RECQ基因之一的突变导致基因组不稳定性，并且在Werner's，Bloom和Rothmund-Thompson综合征中对癌症的易感性升高，以及Werner和Rothmund-Thompson综合症的早衰。因此，RECQ蛋白代表了将基因组稳定性与疾病状态联系起来的范式，以及了解DNA复制，重组和修复之间的联系。这使得对RECQ家族的机制对基础科学和医学很重要。该提案中描述的实验旨在通过解决其在体外和In Viva中的分子作用机制来阐明RECQ蛋白的生物学功能。该方法将生化，高分辨率结构和遗传方法集中在细菌RECQ蛋白上，以描绘支持酶活性的结构 - 功能关系。这些实验包括：（1）确定RECQ家族中保守域的作用（解旋酶，RECQ-CT和HRDC结构域），（2）阐明RECQ的Quaternary和第三结构的阐明，以及（3）Recq-Recq型蛋白质复合物在细菌细胞中的蛋白质和REC研究蛋白质蛋白质蛋白质中的蛋白质复合物的鉴定。一种多学科的方法将允许在RECQ蛋白的细胞作用及其分子作用机理之间建立生化联系。