INITIATION OF LAGGING-STRAND DNA SYNTHESIS

滞后链 DNA 合成的起始

• 批准号: 3285789
• 负责人: KENNETH J MARIANS
• 金额: $ 22.54万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 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 1992-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3285789
• 关键词: DNA binding protein; DNA directed DNA polymerase; DNA footprinting; Escherichia coli; adenosinetriphosphatase; circular DNA; exopeptidase; genetic manipulation; methylation; molecular cloning; nucleic acid sequence; point mutation; proteins; transposon /insertion element

A major part of the effort of this laboratory is directed at developing an understanding of how the replication machinery gains access to the DNA template and subsequently remains bound over tremendous distances while actively synthesizing both the leading- and lagging-strands of DNA. To this end, detailed investigations of the activities of enzymes present at the replication fork are presented. Particular attention is given to the protein-protein and protein-DNA interactions that characterize the replication fork. Studies will be continued that are designed to characterize the interaction of the primosome (a mobile priming apparatus that requires seven E. coli-encoded proteins for assembly and which can assemble at a specific site on the DNA (primosome assembly sites), migrate processively 5' to 3' and occasionally synthesize a primer) with primosome assembly sites. The protein responsible for primosome movement will be established and the three genetically undefined primosomal proteins (replication factor Y (protein n') and proteins n and n") will be molecularly cloned and their genes characterized. The structure and movement of the replication fork will be investigated using novel primer-templates that sustain rolling- circle DNA replication in which synthesis of the leading- and lagging-strands of DNA are coupled. The protein-protein interactions required and the role of the DNA polymerase III holoenzyme in coupling the replication machinery at the fork will be established. Rates of replication fork movement will be measured and factors that influence progress of the replication fork will be identified.

该实验室的主要工作是针对 了解复制机制如何获得优势 进入 DNA 模板并随后保持结合 巨大的距离，同时积极综合两个领先的 和DNA的滞后链。 为此，详细调查 复制叉上存在的酶的活性是 提出。 特别关注蛋白质-蛋白质 以及表征复制的蛋白质-DNA 相互作用 叉。 将继续进行旨在描述特征的研究 引发体（一种移动引发装置， 需要七种大肠杆菌编码的蛋白质进行组装，并且可以 在 DNA 上的特定位点（引发体组装位点）组装， 逐步迁移 5' 至 3'，偶尔合成引物） 具有引发体组装位点。 该蛋白质负责 启动体运动将被建立并且这三个基因 未定义的初体蛋白（复制因子 Y（蛋白 n'） 和蛋白质 n 和 n") 将被分子克隆，它们的基因 特点。 复制叉的结构和运动将是 使用维持滚动的新型底漆模板进行了研究 循环DNA复制，其中前导-和 DNA 的滞后链是耦合的。 蛋白质-蛋白质 所需的相互作用以及 DNA 聚合酶 III 的作用 在叉子处耦合复制机器的全酶将 被成立。 复制叉移动的速率为 测量和影响复制进度的因素 叉将被识别。