FUNCTIONAL ROLES OF CELLULAR PROTEINS IN DNA REPLICATION

细胞蛋白质在 DNA 复制中的功能作用

• 批准号: 6519413
• 负责人: MARC S WOLD
• 金额: $ 30.13万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6519413
• 关键词: DNA binding protein; DNA repair; DNA replication; X ray crystallography; intermolecular interaction; molecular genetics; nuclear magnetic resonance spectroscopy; phosphorylation; protein structure function; yeasts

The human genome is composed of approximately 4x10/9 base pairs of DNA. In order to grow, a human cell must precisely duplicate this entire genome during each cell cycle. This requires the concerted action of many proteins to initiate DNA replication specifically to multiple origins and to synthesize large amounts of DNA accurately. The goal of this research is to elucidate the molecular mechanism(s) of eukaryotic DNA replication and how DNA replication is regulated in the cell. This research focuses on Replication Protein A (RPA), a multi-subunit protein that binds to single-stranded. This human protein is absolutely required for DNA replication, DNA repair and recombination. In addition to binding to single-stranded DNA, RPA interacts with regulation (e.g. p53). These interactions appear to be required for RPA function. RPA also become phosphorylated appears to regulated RPA-protein interactions. The goal of these studies is to elucidate the role of RPA in DNA replication and other reactions of cellular metabolism. A combination of mutational analyses, direct biochemical assays, replication assays and genetic analysis will be used to: (1) define the molecular interactions of RNA with DNA, (2) elucidate the roles of RNA in DNA replication and determine the role(s) of RPA in cellular coordination of DNA metabolism and (3) determine the structure of RPA and characterize conformation changes in RPA. These studies will lead to a better understanding of the molecular mechanism of DNA replication and cellular DNA metabolism.

人类基因组由大约 4x10/9 碱基对的 DNA 组成。为了生长，人类细胞必须在每个细胞周期中精确地复制整个基因组。这需要许多蛋白质的协同作用来启动DNA复制到多个起点并准确地合成大量DNA。本研究的目的是阐明真核 DNA 复制的分子机制以及 DNA 复制在细胞中是如何受到调节的。本研究重点关注复制蛋白 A (RPA)，这是一种与单链结合的多亚基蛋白。这种人类蛋白质是 DNA 复制、DNA 修复和重组所必需的。除了与单链 DNA 结合外，RPA 还与调节（例如 p53）相互作用。这些交互似乎是 RPA 功能所必需的。 RPA 也被磷酸化，似乎可以调节 RPA-蛋白质相互作用。这些研究的目的是阐明 RPA 在 DNA 复制和细胞代谢的其他反应中的作用。突变分析、直接生化分析、复制分析和遗传分析的组合将用于：(1) 定义 RNA 与 DNA 的分子相互作用，(2) 阐明 RNA 在 DNA 复制中的作用并确定其作用RPA 在 DNA 代谢的细胞协调中的作用；(3) 确定 RPA 的结构并表征 RPA 的构象变化。这些研究将有助于更好地了解 DNA 复制和细胞 DNA 代谢的分子机制。