Cell Cycle Regulation of Mcm Localization

MCM 定位的细胞周期调控

• 批准号: 7121153
• 负责人: Muluye E Liku
• 金额: $ 3.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7121153
• 关键词: DNA replication; Saccharomyces cerevisiae; cell cycle; cell growth regulation; cyclin dependent kinase; enzyme activity; fluorescence microscopy; fungal proteins; green fluorescent proteins; natural gene amplification; phosphorylation; predoctoral investigator; protein localization; protein transport; regulatory gene

DESCRIPTION (provided by applicant): We are using the budding yeast Saccharomyces cerevisiae to understand how eukaryotic cells limit DNA replication to one round per cell cycle. Disruption of this regulation can lead to genomic instability, one of the hallmarks of cancer. One conserved mechanism to prevent re-replication is to exclude initiation I proteins from the nucleus after replication initiation has occurred. In budding yeast, nuclear exclusion of the heterohexameric Mcm2-7 replication complex occurs through regulation of Mcm nucleocytoplasmic transport by the cyclin-dependent kinase Cdc28. We have identified nuclear localization signals (NLSs) on Mcm2 and Mcm3 and a potential nuclear export signal (NES) on Mcm3. We have also shown that Cdc28 phosphorylation of Mcm3 near its NLS and NES promotes the net nuclear export of the Mcm2-7 complex. This phosphorylation could work by inhibiting nuclear import, stimulating nuclear export, or doing both. To investigate these possibilities, I will pursue the following specific aims: Aim 1: Determine whether and how Cdc28 regulates Mcm nuclear import Aim 2: Determine whether and how Cdc28 regulates Mcm nuclear export

描述（由申请人提供）：我们正在使用酿酒酵母的萌芽酵母菌来了解真核细胞如何将DNA复制限制为每个细胞周期的一轮。这种调节的破坏会导致基因组不稳定性，这是癌症的标志之一。一种防止重新复制的保守机制是在复制起始后将起始I蛋白排除在核中。在萌芽的酵母中，通过调节细胞周期蛋白依赖性激酶CDC28的MCM核总质胞质转运而发生杂项MCM2-7复制复制复合复制复合复合复制复合复合复合复合复合物的核排除。我们已经确定了MCM2和MCM3上的核定位信号（NLSS）以及MCM3上的潜在核出口信号（NES）。我们还表明，MCM3在其NLS附近和NES附近的Cdc28磷酸化促进了MCM2-7复合物的净核出口。这种磷酸化可以通过抑制核进口，刺激核出口或同时进行。为了调查这些可能性，我将追求以下具体目标： AIM 1：确定CDC28是否以及如何调节MCM核进口 目标2：确定CDC28是否以及如何调节MCM核出口