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CALCIUM, PROTEIN PHOSPHORYLATION, AND CELL CYCLE CONTROL

钙、蛋白质磷酸化和细胞周期控制

基本信息

批准号：
3303136
负责人：
ROGER D SLOBODA
金额：
$ 16.93万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
1990
资助国家：
美国
起止时间：
1990-06-01 至 1993-11-30
项目状态：
已结题

项目摘要

The purpose of the experiments outlined in this application is to provide, from a cell biology point of view, new information concerning the factors controlling the progression of cells through the cell cycle. The many forms of cancer have as one of their common characteristics uncontrolled division of the cancerous cells. Thus the cells divide repeatedly, in amounts and place where they are not required, causing much damage. The experiment proposed here will continue a biochemical and molecular characterization of one protein that is involved in microtubule stability during mitosis and thus may be a key regulator of cell division. Specifically, the experiments proposed are designed to define the characteristics of a recently identified protein (Mrel = 62 kD) that is the substrate for calcium dependent phosphorylation at the start of anaphase. In living cells, a pulse of calcium is released at the metaphase-anaphase transition which is thought to trigger continued transit through the cell cycle. To understand the nature of this trigger more completely, the 62 kD protein will be characterized. To do this, the abundance and distribution of the protein antibodies essential for these experiments have been purified and characteristics will be determined in vitro and in vivo. The former series of experiments will determine the ability of the protein to interact with microtubules in an in vitro assembly system, and test how the protein is involved in microtubule disassembly. For the in vivo experiments, the purified protein will be microinfected into living cells at various times during the cell cycle to test the affect of the protein on normal cell cycle progression. Finally, clones encoding the 62 kD protein will be isolated from a sea urchin expression library to begin molecular analysis of the protein to compare it to other known proteins involved in cell cycle control. The experiments proposed in this application are important because they will provide new and definitive information concerning the control of cell division by beginning an identification of the factors involved in the cascade of events that signal the start of anaphase of mitosis, and, ultimately, the completion of cell division.

本申请中概述的实验的目的是提供，从细胞生物学的角度来看，关于这些因素的新信息通过细胞周期控制细胞的进程。的许多癌症的共同特征之一是不受控制癌细胞的分裂。因此，细胞反复分裂，数量和地方，他们不需要，造成很大的损害。的这里提出的实验将继续一个生物化学和分子一种参与微管稳定性的蛋白质的表征因此可能是细胞分裂的关键调节器。具体而言，所提出的实验旨在定义最近鉴定的蛋白质（Mrel = 62 kD）的特征，在后期开始时钙依赖磷酸化的底物。在活细胞中，钙脉冲在细胞分裂中期-后期释放这种转变被认为会触发细胞的持续转运周期为了更全面地理解这种触发的性质，蛋白质将被表征。为了做到这一点，这些实验所必需的蛋白质抗体将在体外和体内确定纯化和特性。的前一系列的实验将确定蛋白质的能力，在体外组装系统中与微管相互作用，并测试蛋白质参与微管分解。用于体内实验中，纯化的蛋白质将被微感染到活细胞中，在细胞周期的不同时间测试蛋白质对细胞周期的影响正常的细胞周期进程。最后，克隆编码62 kD蛋白将从海胆表达文库中分离，以开始分子分析蛋白质，将其与其他已知的蛋白质进行比较，细胞周期调控本申请中提出的实验是因为它们将提供新的和明确的信息关于控制细胞分裂，事件的级联中所涉及的因素，有丝分裂后期，最终完成细胞分裂。