Genetic and biochemical analysis of cdc-14

cdc-14 的遗传和生化分析

• 批准号: 7900902
• 负责人: R. MAKO SAITO
• 金额: $ 28.49万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-17 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7900902
• 关键词: 3' Untranslated Regions; Animal Model; Be++ element; Beryllium; Biochemical; Biological Models; Caenorhabditis elegans; Cell Cycle; Cell Cycle Regulation; Cell Nucleus; Cell division; Cells; Cues; Cyclin-Dependent Kinase Inhibitor; Cytoplasm; Defect; Development; Elements; Family; Foundations; G1 Phase; Gene Mutation; Gene Order; Genes; Genetic; Genetic Epistasis; Genetic Screening; Goals; Growth and Development function; Homologous Gene; Human; Imagery; Link; Maintenance; Malignant Neoplasms; Mediating; Molecular; Organism; Pathway interactions; Pattern; Phase; Phosphoric Monoester Hydrolases; Play; Process; Production; Relative (related person); Reporter; Reporter Genes; Research; Role; Series; Signal Transduction; Specific qualifier value; Structure; Techniques; Testing; Tissues; Transgenes; Tumor Suppressor Proteins; Yeasts; base; cell type; design; developmental genetics; genome-wide; in vivo; insight; mRNA Stability; member; metaplastic cell transformation; novel; physical property; positional cloning; precursor cell; research study; tumor; yeast two hybrid system

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.

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。建议的研究利用模式生物秀丽线虫来研究发育信号和细胞周期之间的调控相互作用。我们的研究计划建议确定控制CDC-14的途径，CDC-14是发育过程中细胞周期进入的关键调节因子。CDC-14与CKI-1共同作用，CKI-1是细胞周期抑制因子p27Kip1家族中的一员，在调节细胞周期停滞的网络中发挥作用。我们的策略应用线虫优秀的发育遗传学来揭示通过调节CDC-14活性来抑制G1进展的详细机制。 我们提出了三个目标来确定连接发育信号和控制G1进程的途径：1)确定用于限制CDC-14活性的机制，在这些机制中，这种磷酸酶指定了发育过程中暂时的细胞周期停滞。我们集中在两个过程，指导CDC-14活性的细胞类型规范：限制表达和亚细胞区隔。这些分析利用了一系列修饰的CDC-14转基因，旨在操纵CDC-14的表达和亚细胞定位以及这些转基因的相应的G1调节活性。2)在反向遗传筛查中确定在CDC-14介导的网络中作用的其他基因，以寻找控制G1进展的缺陷。我们的屏幕利用一种策略，在细胞周期控制中产生可见的缺陷，从而允许高效和简单的识别。3)使用所描述的屏幕中识别的基因来开发遗传层级。这个网络的组织将基于测试，以确定遗传相互作用，对CDC-14表达的影响，以及新基因的亚细胞定位。总之，拟议的研究将定义在发育过程中控制CDC-14活性的途径，并将提供对用于整合细胞周期调节与生长和发育的一般机制的见解。这项提议的目标是了解用于控制CDC-14活性的机制，CDC-14基因在暂时阻止细胞分裂方面发挥着重要作用。这些结果可能为深入了解肿瘤是如何形成的，因为癌症的特征是控制细胞分裂的基因缺陷。