CELL CYCLE REGULATION DURING ADIPOCYTE DIFFERENTIATION

脂肪细胞分化过程中的细胞周期调控

基本信息

批准号：
2545980
负责人：
DAWN E POST
金额：
$ 2.53万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
1997
资助国家：
美国
起止时间：
1997-09-18 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/2545980
关键词：
3T3 cells adipocytes antisense nucleic acid cell differentiation cell growth regulation cyclins enzyme activity oncoproteins protein kinase

项目摘要

Terminal cell differentiation is accompanied by permanent withdrawal from the cell division cycle. The molecular mechanisms that couple these two processes are poorly understood. Progression of eukaryotic cells through the cell cycle is primarily regulated by a family of structurally related serine/threonine protein kinases, which consists of a regulatory subunit, a cyclin, and a catalytic subunit, a CDK (cyclin dependent kinase). The enzymatic activity of the CDK subunit is regulated by cyclin binding and activation, subunit phosphorylation, and association with CDK inhibitors. The ability of CDK inhibitors to arrest the cell cycle and their induction in response to a variety of growth inhibitory signals, including growth suppression by both the pRb and p53 tumor suppressors, present the CDK inhibitors as a group of ideal molecules that may link cell cycle arrest to cell differentiation. Using the 3T3-L1 preadipocyte cell line as a model system, the subunit composition of CDK complexes, CDK kinase activity, and cell cycle progression during adipocyte differentiation will be analyzed. The function of two CDK inhibitors, p18 and p21, in causing and/or maintaining cell cycle arrest during adipocyte differentiation, as suggested by preliminary experiments, will be tested directly using antisense RNA technology. The involvement of the Rb protein family and p53 in regulating terminal adipocyte differentiation and their interaction with CDK inhibitor pathways will be determined using the HPV16-E6 and HPV16-E7 oncoproteins and dominant negative mutants of pRb and p53.

细胞终末分化伴随着细胞的永久退出细胞分裂周期。将这两者耦合的分子机制对流程了解甚少。真核细胞的进展通过细胞周期主要由结构相关的家族调节丝氨酸/苏氨酸蛋白激酶，由调节亚基组成，细胞周期蛋白和催化亚基 CDK（细胞周期蛋白依赖性激酶）。这 CDK 亚基的酶活性受细胞周期蛋白结合的调节激活、亚基磷酸化以及与 CDK 抑制剂的关联。 CDK 抑制剂阻止细胞周期及其诱导的能力响应各种生长抑制信号，包括生长受到 pRb 和 p53 肿瘤抑制因子的抑制，呈现 CDK 抑制剂作为一组可能与细胞周期停滞相关的理想分子到细胞分化。使用 3T3-L1 前脂肪细胞系作为模型系统，CDK复合物的亚基组成，CDK激酶脂肪细胞分化过程中的活性和细胞周期进展将进行分析。两种 CDK 抑制剂 p18 和 p21 在引起和/或在脂肪细胞分化过程中维持细胞周期停滞，如通过初步实验建议，将直接使用进行测试反义RNA技术。 Rb 蛋白家族的参与和 p53 调节脂肪细胞终末分化及其相互作用 CDK 抑制剂途径将使用 HPV16-E6 和 HPV16-E7 癌蛋白以及 pRb 和 p53 的显性失活突变体。