INITIATION OF DNA REPLICATION AT VIRUS AND CELL ORIGINS

DNA 复制在病毒和细胞起源处的起始

• 批准号: 3481111
• 负责人: BRUCE W. STILLMAN
• 金额: $ 22.29万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-07-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3481111
• 关键词: Adenoviridae; DNA directed DNA polymerase; Escherichia coli; HeLa cells; R factors; affinity chromatography; antibody; binding proteins; cell cycle; circular DNA; genetic manipulation; genetic regulation; human tissue; messenger RNA; nucleic acid chemical synthesis; nucleic acid sequence; nucleobase; simian virus 40; virus DNA; virus genetics; virus protein; virus replication

The mechanism and control of DNA replication in eukaryotic cells is a fundamental problem in understanding cell proliferation. Studies on both virus and cellular DNA replication origins and the proteins that interact with these sequences will be investigated. A cellular protein, nuclear factor I, binds to the origin of human adenovirus DNA replication and is required for the initiation of DNA synthesis. A detailed study on the cell biology of this protein is proposed, specifically to examine the intracellular localization, protein modification and regulation of activity of nuclear factor I. These studies will also involve clonging the gene encoding nuclear factor I and measuring gene expression throughout the cell cycle. Studies on the binding site of this protein, in both virus and cellular DNAs, are proposed. This work will be extended to include similar studies on a cellular protein that binds to a putative origin of replication in the yeast S. cerevisiae. The function of this protein and its DNA binding site in the replication and maintenance of extrachromosomal plasmids and replication of chromosomal DNA will be investigated. Other DNA sites that bind to this cellular protein will be cloned and it will be determined if they have the capability to replicate autonomously in yeast (ARS activity). The gene encoding this protein will be cloned, leading to genetic analysis of the protein's function in cells.

真核细胞中 DNA 复制的机制和控制是 理解细胞增殖的基本问题。 对两者的研究 病毒和细胞 DNA 复制起点以及相互作用的蛋白质 将研究这些序列。 一种细胞蛋白质，核 因子 I，与人类腺病毒 DNA 复制起点结合，是 启动 DNA 合成所需的。 对细胞的详细研究 提出了这种蛋白质的生物学，特别是为了检查 细胞内定位、蛋白质修饰和活性调节 核因子 I。这些研究还将涉及克隆基因 编码核因子 I 并测量整个细胞的基因表达 循环。 对该蛋白质在病毒和病毒中的结合位点的研究 提出了细胞DNA。 这项工作将扩展到包括对细胞蛋白质的类似研究 与酿酒酵母中假定的复制起点结合。 该蛋白及其 DNA 结合位点在复制中的功能 染色体外质粒的维持和染色体的复制 DNA将被调查。 与该细胞结合的其他 DNA 位点 蛋白质将被克隆并确定它们是否具有 在酵母中自主复制的能力（ARS 活性）。 基因 编码该蛋白质的基因将被克隆，从而进行基因分析 蛋白质在细胞中的功能。