MECHANISM AND CONTROL OF INTIATION OF DNA REPLICATION

DNA复制起始的机制和控制

• 批准号: 2166668
• 负责人: ARTHUR KORNBERG
• 金额: $ 74.65万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2166668
• 关键词: DNA directed DNA polymerase; DNA directed RNA polymerase; DNA replication; DNA replication origin; Escherichia coli; adenosine triphosphate; bacterial DNA; bacterial genetics; bacteriophage phi X174; cell cycle; cell growth regulation; cell membrane; electron microscopy; eukaryote; genetic manipulation; helicase; microorganism genetics; molecular genetics; prokaryote; virus genetics

We intend to characterize the numerous enzymes and proteins which we have identified and isolated as participants in the initiation of a cycle of replication of the E. coli chromosome at its origin (oriC) and the initiation of nascent chains at replications forks. These are some of our specific objectives: (i) Described in molecular detail how the origin of the E. coli chromosome is recognized by dnaA protein and so configured to create a bubble for the priming of replication. (ii) Discover how the dnaB helicase, with the aid of dnaC protein, is positioned to create bidirectional forks of replication. (iii) Determine the mechanisms and varieties of transcription activation of initiation at oriC. (iv) Examine the fate and cycling of the proteins that participate in initiation, the influence of membranes, and of ATP and energy charge. (v) Explore existence of factors (positive or negative) which may serve as signals from the increase in cell mass that commit the cell to initiate a new cycle of replication. (vi) Determine the genetic loci and prepare mutants of the primosomal proteins (n, n', and n") to evaluate their physiological importance and their operation at the replication forks of the E. coli chromosome. These biochemical studies are essential for revealing the facts and patterns of replication basic to understanding cell growth and cell division in eukaryotic as well as prokaryotic organisms. With a grasp of the biochemical nature of the switch that operates the initiation of replication, will come opportunities to discover the factors that regulate orderly growth and those responsible for uncontrolled proliferation (e.g. cancer) or its premature cessation.

我们打算表征众多酶和蛋白质 我们已确定并隔离为发起活动的参与者 大肠杆菌染色体在其起源处的复制周期 (oriC) 和复制叉处新生链的起始。 这些是我们的一些具体目标： (i) 从分子角度详细描述了大肠杆菌的起源 染色体被 dnaA 蛋白识别，因此被配置为 创造一个气泡来启动复制。 (ii) 了解 dnaB 解旋酶如何在 dnaC 蛋白的帮助下， 定位于创建双向复制分叉。 (iii) 确定转录的机制和种类 oriC 处起始的激活。 (iv) 检查参与的蛋白质的命运和循环 在起始过程中，膜、ATP 和能量的影响 收费。 (v) 探索可能存在的因素（积极或消极） 作为细胞质量增加的信号， 细胞启动新的复制周期。 (vi) 确定遗传位点并制备突变体 原始体蛋白（n、n' 和 n"）以评估其生理学 重要性及其在 E 复制叉上的操作。 大肠杆菌染色体。 这些生化研究对于揭示事实和 复制模式是理解细胞生长和细胞的基础 真核生物和原核生物的分裂。 与一个 掌握操作开关的生化性质 复制的开始，将会有机会发现 调节有序生长的因素和负责的因素 不受控制的增殖（例如癌症）或其过早 停止。