MECHANISM OF CYCLIN A-CDK2 INHIBITION BY P57

拟议研究的广泛、长期目标是 了解细胞周期调控。 这可能会导致新的战略， 控制与细胞周期相关的健康问题，例如 癌症和生长缺陷。 p57可以通过抑制细胞周期的催化活性， 细胞周期蛋白依赖性激酶（Cdks）。因此，p57作为潜在的负 生长调节剂和候选肿瘤抑制剂。 拟议研究 将利用现代生物物理学和分子生物学技术， 阐明p57抑制cyclin A-Cdk 2的机制。 具体目标是： 1.确定p57抑制的细胞周期蛋白A-Cdk 2复合物的组成 使用沉降平衡。 2.通过以下方法表征p57 Cdk抑制结构域的结构： 核磁共振 3.鉴定p57中参与细胞周期蛋白的氨基酸残基 通过p57的定点诱变抑制A-Cdk 2。 4.鉴定细胞周期蛋白A和/或Cdk 2的氨基酸残基， 通过细胞周期蛋白A和Cdk 2定点突变与p57相互作用。

The broad, lone term objective of the proposed research is to understand cell cycle regulation. This may lead to new strategies for the control of health problems associated with the cell cycle, such as cancer and growth defects. p57 can arrest the cell cycle by inhibiting the catalytic activity of cyclin-dependent kinases (Cdks). p57 thus acts as a potential negative growth regulator and candidate tumor suppressor. The proposed research will use modern biophysical and molecular biology techniques to elucidate the mechanism of cyclin A-Cdk2 inhibition by p57. The specific aims are: 1. Determine the composition of the p57 inhibited cyclin A-Cdk2 complex using sedimentation equilibrium. 2. Characterize the structure of the p57 Cdk-inhibition domain by nuclear magnetic resonance. 3. Identify the amino acid residues of p57 that contribute to cyclin A-Cdk2 inhibition by site-directed mutagenesis of p 57. 4. Identify the amino acid residues of cyclin A and/or Cdk2 that interact with p57 by site-directed mutagenesis of cyclin A and Cdk2.