Genetic and biochemical analysis of cdc-14

描述（由申请人提供）：我们的总体目标是了解调节细胞分裂的分子机制。拟议的研究利用模式生物秀丽隐杆线虫来研究发育信号和细胞周期之间的调节相互作用。我们的研究计划是确定控制cdc-14的途径，cdc-14是发育过程中进入细胞周期的关键调节因子。cdc-14与细胞周期蛋白依赖性激酶抑制剂p27 Kip 1家族的成员cki-1一起在调节细胞周期静止的网络中起作用。我们的策略应用了C. elegans揭示通过调节cdc-14活性来抑制G1进展的详细机制。 我们提出了三个目标，以确定连接发育信号控制G1进程的途径：1）确定用于限制cdc-14活性的机制，在这些细胞中，这种磷酸酶在发育过程中暂时指定细胞周期静止。我们集中在两个过程，直接cdc-14活性的细胞类型规范：限制性表达和亚细胞区室化。这些分析利用一系列经修饰的cdc-14转基因，其设计用于操纵CDC-14表达和亚细胞定位以及此类转基因的相应G1调节活性。2)鉴定在cdc-14介导的网络中起作用的其他基因，以进行反向遗传筛查，以控制G1进展的缺陷。我们的屏幕利用一种策略，以产生可见的缺陷，在细胞周期控制，允许有效和简单的识别。3)使用所述筛选中鉴定的基因建立遗传层次。该网络的组织将基于确定新基因的遗传相互作用、对CDC-14表达的影响和亚细胞定位的测试。总之，这些研究将确定发育过程中控制cdc-14活性的途径，并将提供对细胞周期调控与生长发育相结合的一般机制的见解。这项计划的目标是了解cdc-14基因的活性控制机制，cdc-14基因在暂时阻止细胞分裂中起着重要作用。这些结果可能会提供关于肿瘤如何形成的见解，因为癌症的特征是控制细胞分裂的基因缺陷。

DESCRIPTION (provided by applicant): Our overall goal is to achieve an understanding of the molecular mechanisms regulating cell divisions. The proposed studies utilize the model organism Caenorhabditis elegans to investigate the regulatory interactions between developmental signals and the cell cycle. Our research plan proposes to identify the pathways that control cdc-14, a critical regulator of cell-cycle entry during development. cdc-14 acts together with cki-1, a member of the p27Kip1 family of cyclin-dependent kinase inhibitors, within a network that regulates cell-cycle quiescence. Our strategies apply the excellent developmental genetics of C. elegans to reveal detailed mechanisms to inhibit G1 progression by modulating cdc-14 activity. We propose three aims to identify the pathways that connect developmental signals to control of G1 progression: 1) To determine the mechanisms used to restrict cdc-14 activity to those cells in which this phosphatase specifies temporarily cell-cycle quiescence during development. We focus on two processes that direct the cell-type specification of cdc-14 activity: restricted expression and subcellular compartmentalization. These analyses utilize a series of modified cdc-14 transgenes designed to manipulate CDC-14 expression and subcellular localization and the corresponding G1 regulatory activity of such transgenes. 2) To identify additional genes that act within the cdc-14-mediated network in a reverse-genetic screen for defects in control of G1 progression. Our screen utilizes a strategy to produce visible defects in cell- cycle control, allowing for efficient and simple identification. 3) To develop a genetic hierarchy using the genes identified in the described screen. The organization of this network will be based on tests to define the genetic interactions, effect on CDC-14 expression and subcellular localization of the new genes. Together, the proposed studies will define the pathway to control cdc-14 activity during development and will provide insights into the general mechanisms used to integrate cell-cycle regulation with growth and development. The goal of this proposal is to understand the mechanisms used to control the activity of cdc-14, a gene that plays an important role to temporarily stop cells from dividing. These results may provide insights into how tumors are formed since cancers are characterized by defects in genes that control cell divisions.