INITIATION OF DNA REPLICATION OF YEAST CHROMOSOMES

酵母染色体 DNA 复制的起始

• 批准号: 6636120
• 负责人: Stephen P. Bell
• 金额: $ 25.96万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6636120
• 关键词: DNA; DNA binding protein; DNA replication; DNA replication origin; Saccharomyces cerevisiae; adenosine triphosphate; cell cycle; cell cycle proteins; chromatin; chromosomes; conformation; crosslink; electron microscopy; fungal genetics; fungal proteins; gene deletion mutation; laboratory mouse; molecular cloning; monoclonal antibody; nucleosomes; protein protein interaction; protein structure function; proteolysis; tissue /cell culture; yeasts

DESCRIPTION: The timely and accurate replication of the genome is essential to the normal proliferation of all eukaryotic cells. Accordingly the initiation of DNA replication is carefully coordinated with the progress of the cell cycle. The long-term objective of this proposal is to determine the events that select the sites of DNA replication and activate the initiation process. The origin recognition complex (ORC) is a key factor controlling these events. ORC is the only S. cerevisiae protein known to specifically recognize all yeast origins of replication. Once bound to these sites, ORC acts to recruit other replication factors to the origin of replication. In this proposal, Dr. Bell will exploit his recent advances in understanding the biochemical functions of ORC to determine how these functions are mechanistically coupled to the replication initiation process. Specifically Dr. Bell will: Determine the mechanism of DNA binding by ORC and how this event alters the function of ORC in the cell. These studies will provide critical information concerning the mechanisms that select origins of replication. Determine the function of ORC during replication complex assembly and activation at origins of replication. Using both biochemical analysis of replication factor interaction and a genetic screen for novel ORC mutants, these studies will define key events required for the assembly of essential replication complexes. Determine the influence of nucleosomes on ORC and origin function. Using nucleosome assembly reactions and a novel genome-wide assay for determining the sites of ORC binding, Dr. Bell will dissect the impact of nucleosomes and chromatin on DNA replication. The highly conserved nature of eukaryotic DNA replication strongly argues that any new understanding of the fundamental mechanisms of eukaryotic DNA replication in yeast will be readily translated to our understanding of the same process in human cells. The rapid progress that can be made in S. cerevisiae make it an ideal organism to perform these studies. New understanding of these proteins will lead to strong candidate targets for anti-fungal compounds and potential markers to identify cancer cells. In addition, the understanding of the yeast replication proteins gained in these studies will direct studies to identify inhibitors of the human analogs of these proteins, which represent important targets for novel chemotherapeutic compounds.

描述：基因组的及时，准确复制对于 所有真核细胞的正常增殖。因此 DNA复制与细胞周期的进展仔细协调。 该提议的长期目标是确定选择的事件 DNA复制的位点并激活启动过程。起源 识别复合物（ORC）是控制这些事件的关键因素。兽人是 只有酿酒酵母蛋白已知才能特异性地识别 复制。一旦绑定到这些站点，兽人就可以招募其他复制 复制起源的因素。在此提案中，贝尔博士将利用 他最近了解ORC的生化功能的进步 确定这些功能如何机械地耦合到复制 启动过程。特别是贝尔博士将：确定DNA的机制 ORC结合以及此事件如何改变ORC在单元格中的功能。这些 研究将提供有关选择机制的关键信息 复制的起源。在复制过程中确定兽人的功能 复杂的复杂组装和激活。两者都使用 复制因子相互作用和遗传筛查的生化分析 新型的兽人突变体，这些研究将定义 基本复制复合物的组装。确定 兽人和原点功能上的核小组。使用核小体组装反应和 贝尔博士，一种用于确定兽人结合位点的新型基因组测定 将剖析核小体和染色质对DNA复制的影响。这 真核DNA复制的高度保守性强烈认为任何 对真核DNA复制的基本机制的新理解 在酵母中，将很容易转化为我们对同一过程的理解 人类细胞。可以在酿酒酵母中取得的快速进步使它成为 理想的有机体进行这些研究。对这些蛋白质的新理解 将导致强大的抗真菌化合物和潜力的候选目标 标记以鉴定癌细胞。此外，对酵母的理解 这些研究中获得的复制蛋白将指导研究以识别 这些蛋白质的人类类似物的抑制剂，这些蛋白质代表重要 新型化学治疗化合物的靶标。