MECHANISMS OF ESCHERICHIA COLI CHROMOSOMAL REPLICATION

大肠杆菌染色体复制机制

• 批准号: 3284293
• 负责人: JON M KAGUNI
• 金额: $ 13.1万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3284293
• 关键词: DNA binding protein; DNA directed RNA polymerase; Escherichia coli; adenosine triphosphate; autoradiography; bacterial genetics; bacterial proteins; binding proteins; cell growth regulation; enzyme linked immunosorbent assay; enzyme mechanism; gel electrophoresis; gene expression; gene interaction; genetic manipulation; genetic mapping; genetic promoter element; genetic strain; genetic transcription; high performance liquid chromatography; hydrolysis; immunoelectron microscopy; laboratory rabbit; molecular cloning; mutant; nucleic acid hybridization; nucleic acid sequence; plasmids; protein sequence; radioimmunoassay; radiotracer; transposon /insertion element; ultracentrifugation

Chromosomal replication in the bacterium Escherichia coli is strictly coordinated to the growth rate of the bacterium. This coordination is controlled through events which regulate the frequency of initiation of each cycle of replication. The long range objective of this proposed research is to understand biochemically how initiation of chromosomal replication occurs, and how it is regulated and coupled to other cellular processes. One approach of the proposed research involves biochemical characterization of mutant forms of dnaA protein to identify the biochemical alterations which make these mutationally altered proteins unable to function in initiation of replication. The second interelated approach involves biochemical characterization of gene products encoded by extragenic suppressors of dnaA mutants in order to understand the mechanism of suppression. These studies will lend insight into the mechanism of initiation of DNA replication, the function of dnaA protein in initiation, and other factors which interact with dnaA protein and contribute to the initiation of replication. A biochemical understanding of the initiation process and its regulation will provide a framework for how these processes may occur in higher organisms, what events may induce the malignant growth of normally quiescent cells, and how these abnormal processes may be controlled or prevented.

大肠杆菌中的染色体复制是 与细菌的生长速度严格协调。 这 协调是通过调节事件来控制的 每个复制周期的起始频率。 长的 这项研究的目标是了解 生物化学上染色体复制的起始是如何发生的， 以及它如何被调节并与其他细胞过程耦合。 拟议研究的一种方法涉及生物化学 dnaA 蛋白突变形式的表征，以鉴定 使这些发生突变的生化改变 蛋白质无法在复制起始过程中发挥作用。 这 第二个相关方法涉及生物化学 外源编码基因产物的表征 dnaA 突变体的抑制子，以了解 抑制机制。 这些研究将有助于深入了解 DNA复制起始机制，dnaA的功能 起始蛋白以及与 dnaA 相互作用的其他因素 蛋白质并有助于复制的启动。 对起始过程及其影响的生化理解 监管将为这些流程如何提供一个框架 发生在高等生物中，哪些事件可能诱发恶性 正常静止细胞的生长，以及这些异常细胞如何生长 过程可以被控制或阻止。