Cell Cycle control by cyclin-dependent kinases

超出所提供的空间。细胞周期蛋白依赖激酶（Cdks）驱动真核细胞周期的事件。Cdks被认为是通过磷酸化细胞中的大量蛋白质底物来起作用，但这些底物很少被鉴定出来。在拟议的研究中，一种新的生化策略将用于在出芽酵母和脊椎动物中进行大规模的Cdk底物搜索。我们的初步研究已经在出芽酵母中鉴定了大约200个Cdkl底物，在本文的第一个目标中，我们将详细分析这些底物的一个子集。其他的初步工作已经揭示了相关细胞周期蛋白亚基的身份对Cdkl磷酸化许多酵母底物的速率有深远的影响，而拟议研究的第二个目的将是探索这种细胞周期蛋白特异性的分子基础及其在细胞中的生理功能。研究人员还将开发方法，对脊椎动物细胞中大量Cdk底物进行系统鉴定和表征。最后，将采用“化学遗传学”方法构建小鼠和小鼠细胞系，其中Cdkl或Cdk2可以高特异性地抑制，从而可以严格分析它们在正常细胞生物学和肿瘤中的功能。这些研究有望为Cdks驱动细胞周期事件的基本机制提供广泛的新见解。这些见解是朝着更好地理解染色体分离缺陷和癌细胞特征的其他细胞周期缺陷迈出的重要一步。网站性能 ======================================== 节结束 ===========================================

EXCEED THE SPACE PROVIDED. Cyclin-dependent kinases (Cdks) drive the events of the eukaryotic cell cycle. Cdks are thought to act by phosphorylating a large number of protein substrates in the cell, but few of these substrates have been identified. In the proposed studies, a novel biochemical strategy will be used to perform large scale searches for Cdk substrates in budding yeast and vertebrates. Our preliminary studies have led to the identification of about 200 Cdkl substrates in budding yeast, and in the first aim of the proposed work a subset of these substrates will be analyzed in detail. Other preliminary work has revealed that the identity of the associated cyclin subunit has a profound effect on the rate at which Cdkl phosphorylates many yeast substrates, and the second aim of the proposed studies will be to explore the molecular basis of this cyclin specificity and its physiological function in the cell. Methods will also be developed for the systematic identification and characterization of large numbers of Cdk substrates in vertebrate cells. Finally, a 'chemical genetic' approach will be employed to construct mice and mouse cell lines in which Cdkl or Cdk2 can be inhibited with high specificity, allowing rigorous analysis of their function in normal cell biology and in tumors. These studies promise to provide a wide range of new insights into the basic mechanisms by which Cdks drive cell-cycle events. Such insights are an important step toward a better understanding of the chromosome segregation defects and other cell-cycle defects that characterize the cancer cell. PERFORMANCE SITE ========================================Section End===========================================