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 DNA复制的潜在调控起始。 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影响 DNA 和蛋白质-蛋白质相互作用。 细菌质粒提供 研究这一问题的最先进、最方便的系统之一 基本的生物过程。 我们开发了遗传和 生化工具是使用的绝佳切入点 质粒 R6K 作为成分分析的模型系统，以及 确保受控复制的事件 原核基因组。 第一节中提出的实验针对 R6K复制区域关键域的识别 这对于形成涉及的高阶结构至关重要 多功能复制蛋白，PI。 第二部分的实验将利用 从改变的突变体中纯化的 PI 蛋白变体的可用性 在复制起始的阴性对照中探测其作用 负控制回路中的PI。 在第三部分中，我们建议对E的作用进行研究。 质粒 R6K 中的大肠杆菌整合宿主因子 (IHF) 复制。 我们的初步数据使我们相信 IHF 蛋白质可能在细胞结构中发挥关键作用 质粒起源。 由于一大群原核和真核染色体 在组织结构上表现出显着的结构相似性 一个基本的复制子，我们相信通过研究质粒 R6K，我们 最终将了解基本机制 一般而言，这是遗传物质复制的基础。