ROLE OF NUCLEOPROTEIN STRUCTURES IN GENOME DUPLICATION

核蛋白结构在基因组复制中的作用

• 批准号: 3297740
• 负责人: MARCIN S FILUTOWICZ
• 金额: $ 13.92万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-03-01 至 1992-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3297740
• 关键词: DNA binding protein; DNA footprinting; DNA replication; DNA replication origin; Escherichia coli; genetic mapping; genetic regulation; mutant; nucleoproteins; plasmids; protein structure function

Our major objective is to understand the molecular mechanism underlying regulated initiation of DNA replication. DNA replication, like other high precision DNA transactions such as recombination and transcription is largely influenced by protein- DNA and protein-protein interactions. Bacterial plasmids provide one of the most advanced and convenient systems to study this fundamental biological process. We have developed genetic and biochemical tools which are and excellent point of entry to use plasmid R6K as a model system in the analysis of components, and the events that assure the controlled duplication of a prokaryotic genome. The experiments proposed in the first section are directed toward the identification of key domains in the R6K replication region which are critical in forming higher order structures involving the multifunctional replication protein, PI. The experiments in the second section will take advantage of the availability of PI protein variants purified from mutants altered in a negative control of replication initiation to probe the role of PI in negative control loop. In the third section, we propose studies on the role of the E. coli Integration Host Factor (IHF) in the plasmid R6K replication. Our preliminary data convince us that the IHF protein could play a critical role in the architecture of the plasmid origin. Since a large group of prokaryotic and eukaryotic chromosomes exhibit remarkable structural similarities in the organization of a basic replicon, we believe that by studying plasmid R6K, we will ultimately learn about fundamental mechanisms that are underlying the duplication of genetic material in general.

我们的主要目标是了解分子机制 潜在的DNA复制的调控启动。脱氧核糖核酸 复制，就像其他高精度DNA交易一样，例如 重组和转录很大程度上受蛋白质的影响- DNA和蛋白质之间的相互作用。细菌质粒提供了 最先进和最方便的系统之一来研究这个 基本的生物过程。我们已经开发出了基因和 生化工具是使用的绝佳切入点 以R6K为模型体系进行成分分析，以及 确保受控复制 原核生物基因组。 第一部分中提出的实验是针对 R6K复制区关键结构域的鉴定 它们在形成更高层次的结构中起着关键作用 多功能复制蛋白PI。 第二部分中的实验将利用 从突变株中纯化的PI蛋白变异体的可用性 在负调控复制启动中的探测作用 在负控制回路中的PI。 在第三部分中，我们提出了对E。 表达载体R6K中的整合宿主因子(IHF) 复制。我们的初步数据让我们相信IHF 蛋白质可能在生物体的结构中起到关键作用 质粒源。 由于一大群原核和真核染色体 在组织结构上表现出显著的相似性 一个基本的复制子，我们相信通过研究R6K质粒，我们 最终将了解以下基本机制： 通常情况下，遗传物质的复制是潜在的。