MCM PROTEIN FUNCTION DURING EUKARYOTIC DNA REPLICATION

真核 DNA 复制过程中的 MCM 蛋白功能

• 批准号: 6351274
• 负责人: Stephen P. Bell
• 金额: $ 17.96万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6351274
• 关键词: DNA binding protein; DNA replication; Saccharomyces cerevisiae; adenosine triphosphate; cell cycle; fungal genetics; fungal proteins; gene mutation; laboratory mouse; molecular cloning; nucleic acid sequence; protein structure function; site directed mutagenesis; tissue /cell culture

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 direct the initiation of replication and how these factors then contribute to the elongation phases of DNA replication. The MCM proteins are a family of six related proteins that play a central role in the replication process in the yeast S. cerevisiae. Recent studies indicate that these proteins are among the earliest factors assembled at origins of replication (prior to the initiation of DNA replication) but also participate in the elongation phase of the replication process. These and other findings suggest that the MCM proteins act as the replicative DNA helicase in eukaryotic cells. This hypothesis is tested in this proposal by addressing the genetic and biochemical properties of the MCM proteins. Specifically Dr. Bell will: Identify conditional mutants in MCM genes that distinguish between the initiation and elongation functions of these proteins. The resulting mutants will be characterized for their effects on the association of the MCM and other proteins with origin and non-origin DNA sequences. Determine the requirement for origin DNA unwinding to assemble MCM proteins at the origin and identify similar elements in other origins of replication. Determine the biochemical properties of purified MCM proteins with particular attention on their ability to bind and hydrolyze ATP and to act as a DNA helicase. Based on previous studies, 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 a ideal organism to perform these studies. New understanding of these proteins will lead to strong candidate targets for novel anti-fungal compounds. In addition, the understanding of the yeast MCM proteins gained in these studies will direct studies to identify inhibitors of the human analogs of these proteins, which are strong candidate targets for novel chemotherapeutic compounds based on the success of previous DNA replication inhibitors in chemotherapeutic regimens.

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复制的启动与进度仔细协调 细胞周期。 该提议的长期目标是 确定指导重复开始的事件以及如何 这些因素随后有助于DNA的伸长阶段 复制。 MCM蛋白是一个六个相关蛋白质的家族， 在酵母中的复制过程中起着核心作用。 酿酒酵母。 最近的研究表明，这些蛋白质是 最早的因素是在复制的起源（之前） 启动DNA复制），但也参与伸长率 复制过程的阶段。 这些和其他发现表明 MCM蛋白充当真核的复制DNA解旋酶 细胞。 该假设通过解决 MCM蛋白的遗传和生化特性。 具体来说 贝尔博士威尔： 在MCM基因中识别有条件突变体的区分 这些蛋白质的起始和伸长功能。 结果 突变体的特征是它们对关联的影响 具有起源和非原素DNA序列的MCM和其他蛋白质。 确定原点DNA放弃组装MCM的要求 蛋白质处于起源并识别其他起源中的类似元素 复制。 确定具有纯化MCM蛋白的生化特性 特别关注它们结合和水解ATP的能力以及 充当DNA解旋酶。 基于以前的研究，对基本的任何新理解 酵母中真核DNA复制的机制很容易 转化为我们对人类细胞中相同过程的理解。这 可以在S. cerevisiae中取得的快速进步使其成为理想 有机体进行这些研究。对这些蛋白质的新理解 将导致新型抗真菌化合物的强大候选靶标。 此外，对酵母MCM蛋白的理解获得了 这些研究将指导研究以鉴定人类的抑制剂 这些蛋白质的类似物，它们是新颖的强大候选靶标 基于先前DNA成功的化学治疗化合物 化学治疗方案中的复制抑制剂。