MCM PROTEIN FUNCTION DURING EUKARYOTIC DNA REPLICATION

真核 DNA 复制过程中的 MCM 蛋白功能

• 批准号: 6498795
• 负责人: Stephen P. Bell
• 金额: $ 18.47万
• 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 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6498795
• 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.

及时准确地复制基因组对人类的生存至关重要。 所有真核细胞的正常增殖。 因此 DNA复制的启动与DNA复制的进展密切相关。 细胞周期。 这项建议的长远目标是 确定指导复制启动的事件以及如何 这些因素然后有助于DNA的延伸阶段 复制的MCM蛋白是一个由六种相关蛋白质组成的家族， 在酵母S. 啤酒。 最近的研究表明，这些蛋白质是 最早在复制起点组装的因子（在 启动DNA复制），但也参与延伸 复制过程的一个阶段。 这些和其他研究结果表明， MCM蛋白在真核生物中充当复制DNA解旋酶， 细胞 本提案通过解决以下问题来检验这一假设： MCM蛋白的遗传和生物化学性质。 具体 博士贝尔将： 鉴定MCM基因中的条件突变体， 这些蛋白质的起始和延伸功能。 所得 突变体将被表征为它们对以下关联的影响： MCM和其它具有起源和非起源DNA序列的蛋白质。 确定组装MCM所需的起始DNA解旋 蛋白质的起源，并确定在其他来源的类似元素 复制。 测定纯化的MCM蛋白的生化性质， 特别注意它们结合和水解ATP的能力， 作为DNA解旋酶。 基于以前的研究，任何对基本的新的理解， 酵母中真核DNA复制的机制将很容易 转化为我们对人类细胞中相同过程的理解。的 快速的进步，可以在S。酿酒使它成为一个理想的 生物来进行这些研究。对这些蛋白质的新认识 将为新型抗真菌化合物提供强有力的候选靶点。 此外，对酵母MCM蛋白质的了解也在 这些研究将指导研究，以确定人类的抑制剂， 这些蛋白质的类似物，其是新的免疫抑制剂的强候选靶标。 化疗化合物的基础上成功的DNA 化疗方案中的复制抑制剂。

项目成果

Visualization of replication initiation and elongation in Drosophila.

果蝇中复制启动和伸长的可视化。

• DOI: 10.1083/jcb.200207046
• 发表时间: 2002-10-28
• 期刊: JOURNAL OF CELL BIOLOGY
• 影响因子: 7.8
• 作者: Claycomb, Julie M;MacAlpine, David M;Evans, James G;Bell, Stephen P;Orr-Weaver, Terry L
• 通讯作者: Orr-Weaver, Terry L