The role of Cdc27 in miotic regulation of the APC/C

Cdc27 在 APC/C 缩瞳调节中的作用

• 批准号: 6886604
• 负责人: Dina Priscilla Matheos
• 金额: $ 2.3万
• 依托单位: RESEARCH INST OF FOX CHASE CAN CTR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6886604
• 关键词: HeLa cells; Saccharomyces cerevisiae; antimitotics; cell cycle; cell cycle proteins; chromosome movement; postdoctoral investigator

DESCRIPTION (provided by applicant): Chromosome segregation is one of the most critical processes common to every organism. To ensure proper segregation, the cell employs a checkpoint mechanism to monitor defects in the segregation machinery. The mitotic checkpoint ensures that even a single improperly attached chromosome is capable of delaying the metaphase to anaphase transition. To achieve a delay prior to chromosome segregation, the checkpoint inhibits, through a poorly understood mechanism, the anaphase-promoting complex/cyclosome (APC/C). Our preliminary studies revealed that injection of antibodies to the c-terminus of Cdc27, a subunit of the APC/C, abrogated the mitotic checkpoint in HeLa cells. This is supported by preliminary studies in yeast which revealed a c-terminal deletion of Cdc27 that prematurely separates its sister chromatids in the presence of nocodazole. To extend these observations, I propose to use both genetic and biochemical approaches in the budding yeast Saccharomyces cerevisiae and in a mammalian tissue culture system to characterize the interaction between Cdc27 and the mitotic checkpoint and test how this interaction might facilitate mitotic checkpoint inhibition of the APC/C.

描述（由申请方提供）：染色体分离是每种微生物常见的最关键过程之一。为了确保适当的分离，细胞采用检查点机制来监测分离机制中的缺陷。有丝分裂检查点确保即使是单个不正确连接的染色体也能够延迟中期到后期的转变。为了在染色体分离之前实现延迟，检查点通过一种知之甚少的机制抑制后期促进复合物/细胞周期体（APC/C）。我们的初步研究表明，注射抗体到Cdc 27的C-末端，APC/C的亚基，废除了HeLa细胞中的有丝分裂检查点。这得到了酵母中的初步研究的支持，该研究揭示了Cdc 27的C-末端缺失，其在诺考达唑存在下过早地分离其姐妹染色单体。为了扩展这些观察结果，我建议在芽殖酵母酿酒酵母和哺乳动物组织培养系统中使用遗传和生化方法来表征Cdc 27和有丝分裂检查点之间的相互作用，并测试这种相互作用如何促进APC/C的有丝分裂检查点抑制。