Regulatory Enzymes and Systems in Cell Cycle Control

细胞周期控制中的调节酶和系统

• 批准号: 9918408
• 负责人: DAVID Owen MORGAN
• 金额: $ 94.72万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2021-06-08
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918408
• 关键词: Activator Appliances; Award; Behavior; Binding; Biochemical; Biological Process; Cancer Etiology; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell division; Cells; Cellular biology; Chromosome Segregation; Chromosomes; Computer Models; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinases; Cytology; Defect; Development; Disease; Enzymes; Eukaryotic Cell; Event; Genetic; Malignant Neoplasms; Methods; Mitosis; Mitotic; Modification; Phosphoric Monoester Hydrolases; Phosphorylation; Process; Proteins; Regulation; Reproduction; Research; Research Personnel; Saccharomyces cerevisiae; Saccharomycetales; Substrate Interaction; System; Time; Ubiquitination; Work; anaphase-promoting complex; daughter cell; event cycle; genetic regulatory protein; human disease; insight; public health relevance; single molecule; tumor progression; ubiquitin ligase

 DESCRIPTION (provided by applicant): The proposed work explores the regulatory system that controls the eukaryotic cell division cycle, with an emphasis on the mechanisms that trigger cell-cycle events at the correct time and in the correct order, resulting in robust and accurate reproduction of the cell. The cell-cycle control system is composed of master regulatory enzymes that include the cyclin-dependent protein kinases and a ubiquitin ligase called the anaphase-promoting complex or APC. In the proposed studies, these enzymes and their biological functions will be analyzed using biochemical, cytological, and computational approaches in the budding yeast Saccharomyces cerevisiae. Much of the work will focus on mechanisms of APC activation and substrate binding, which depend on transient association of the APC with a rate-limiting activator subunit that binds the substrate. A previously unknown activity that regulates binding of activator to the APC will be identified and characterized, and single-molecule methods will be developed for analysis of APC-activator interactions. Computational modeling will be used to explore how the modulation of APC-substrate interactions leads to the ordered ubiquitination of APC targets during mitosis. Biochemical approaches will be used to dissect the effects of APC phosphorylation on activator function and to explore activator regulation in the spindle assembly checkpoint. The proposed work also includes studies of the regulation of cell-cycle progression by protein phosphorylation, with an emphasis on the control of late mitotic events by a specific phosphatase. The information gained from these studies will provide important new insights into the control of cell-cycle progression, and will thereby enhance our understanding of diseases, such as cancer, in which cell-cycle control or chromosome segregation is defective. These studies will also illuminate general mechanisms of protein ubiquitination and phosphorylation, regulatory modifications of major importance throughout cell biology and human disease.

 描述（由申请人提供）：拟议的工作探索了控制真核细胞分裂周期的调控系统，重点是在正确的时间和正确的顺序触发细胞周期事件的机制，从而导致细胞的稳健和准确的繁殖。细胞周期控制系统由主要的调节酶组成，包括细胞周期蛋白依赖性蛋白激酶和称为后期促进复合物或APC的泛素连接酶。在拟议的研究中，这些酶及其生物学功能将使用生化，细胞学和计算方法在芽殖酵母酿酒酵母中进行分析。大部分工作将集中在APC激活和底物结合的机制，这取决于APC与结合底物的限速激活子亚基的瞬时关联。一个以前未知的活动，调节激活剂的APC结合将被确定和表征，并将开发单分子方法用于分析APC-激活剂相互作用。计算建模将被用来探索APC-底物相互作用的调制如何导致APC靶点在有丝分裂过程中的有序泛素化。生物化学的方法将被用来解剖APC磷酸化对激活剂功能的影响，并探讨激活剂在纺锤体组装检查点的调节。拟议的工作还包括蛋白质磷酸化调节细胞周期进程的研究，重点是通过特定的磷酸酶控制晚期有丝分裂事件。从这些研究中获得的信息将为细胞周期进程的控制提供重要的新见解，从而增强我们对疾病的理解，如癌症，其中细胞周期控制或染色体分离是有缺陷的。这些研究还将阐明蛋白质泛素化和磷酸化的一般机制，在整个细胞生物学和人类疾病中具有重要意义的调节修饰。