Regulation of DNA Replication and Repair

DNA 复制和修复的调控

• 批准号: 7048635
• 负责人: MARK D. SUTTON
• 金额: $ 27.06万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 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/7048635
• 关键词: DNA damage; DNA directed DNA polymerase; DNA repair; DNA replication; DNA replication origin; Escherichia coli; cell cycle; enzyme mechanism; mutant; protein binding

DESCRIPTION (provided by applicant): The long-term goal of this research is to understand how organisms accurately duplicate their genetic material by coordinating the actions of their DNA replication machinery with those of other cellular proteins involved in repair, damage tolerance, and cell cycle progression. Failure of an organism to do so can have catastrophic consequences ranging from disease, such as cancer, to death. The proposed research program utilizes both biochemical and genetic approaches to understand the roles played by the beta processivity clamp of the E. coli replicative DNA polymerase in coordinating DNA replication, repair, damage tolerance, and cell cycle progression. The beta clamp participates in a DNA damage checkpoint control as well as in translesion DNA synthesis (TLS). In addition, it interacts with a variety of proteins known to function in various aspects of DNA metabolism. As one Aim, we will test our hypothesis that unique interactions of beta with the different umuDC gene products influence the choice between replication, checkpoint, and TLS. These studies will have broad relevance to understanding integration of replication and TLS in both prokaryotic and eukaryotic cells. As a second Aim, we will test our proposal that beta participates in at least one DNA repair and/or umuDC-independent damage tolerance function by determining the molecular basis of the UV light sensitivity of a dnaN59 beta mutant. These studies will further our understanding of mechanisms that couple replication and repair. As a third Aim, we will test our hypothesis that certain partner proteins bind to overlapping surfaces on beta, and that by competing with each other for binding to the clamp, beta is able to regulate which proteins gain access to the replication fork. These studies will provide important insights into fundamental mechanisms of polymerase switching and replication fork management. In summation, we anticipate that the study of the coordinated regulation of DNA replication, DNA damage tolerance and repair, and cell cycle progression in E. coli will provide a valuable framework for understanding these same processes in humans, where the complexity of the events is far greater.

描述（由申请人提供）：这项研究的长期目标是通过协调其DNA复制机械的作用与其他参与修复，损伤耐受性和细胞周期进展的细胞蛋白的作用来了解生物如何准确地复制其遗传材料。生物体失败会产生灾难性的后果，从癌症等疾病到死亡。提出的研究计划利用生化和遗传方法来了解大肠杆菌复制DNA聚合酶在协调DNA复制，修复，损伤耐受性和细胞周期进程中所起的作用。 Beta夹参与DNA损伤检查点控制以及Translesion DNA合成（TLS）。此外，它与已知在DNA代谢的各个方面起作用的多种蛋白质相互作用。作为一个目的，我们将测试我们的假设，即β与不同UMUDC基因产品的独特相互作用会影响复制，检查点和TLS之间的选择。这些研究将与理解原核生物和真核细胞中复制和TLS的整合具有广泛的相关性。作为第二个目的，我们将通过确定DNAN59β突变体的UV光敏感性的分子基础来测试Beta参与至少一个DNA修复和/或UMUDC独立损伤耐受函数的建议。这些研究将进一步理解对复制和修复的机制。作为第三个目的，我们将检验我们的假设，即某些伴侣蛋白与Beta上的重叠表面结合，并且通过相互竞争以与夹具结合，beta可以调节哪些蛋白可以访问复制叉。这些研究将为聚合酶切换和复制叉管理的基本机制提供重要的见解。总而言之，我们预计，对DNA复制，DNA损伤耐受性和修复的协调调节以及大肠杆菌中的细胞周期进程将为理解人类中这些相同过程的宝贵框架，其中事件的复杂性要大得多。