MECHANISM OF CYCLIN A-CDK2 INHIBITION BY P57

拟议研究的广泛的、唯一的长期目标是 了解细胞周期调节。这可能会导致新的战略 控制与细胞周期有关的健康问题，如 癌症和生长缺陷。 P57可以通过抑制细胞周期蛋白的催化活性来阻断细胞周期。 细胞周期蛋白依赖性蛋白激酶(CDK)。因此，P57起到了潜在的否定作用 生长调节剂和候选肿瘤抑制因子。拟议的研究 将使用现代生物物理和分子生物学技术来 阐明p57抑制细胞周期蛋白A-CDK2表达的机制。 具体目标是： 1.测定p57抑制的细胞周期蛋白A-CDK2复合体的组成 采用沉降法平衡。 2.表征p57CDK-抑制结构域的结构 核磁共振。 3.确定p57中与细胞周期蛋白有关的氨基酸残基 P57基因定点突变对a-cdk2的抑制作用。 4.鉴定Cyclin A和/或CDK2的氨基酸残基 通过细胞周期蛋白A和CDK2的定点突变与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.