Genetic and biochemical analysis of cdc-14

cdc-14 的遗传和生化分析

• 批准号: 7496963
• 负责人: R. MAKO SAITO
• 金额: $ 28.78万
• 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/7496963
• 关键词: 3' Untranslated Regions; Animal Model; Be++ element; Beryllium; Biochemical Genetics; 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; Genome; Goals; Green Fluorescent Proteins; 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; in vivo; insight; mRNA Stability; member; metaplastic cell transformation; novel; p27 Cell Cycle Protein; p27 Enzyme Inhibitor; 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。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。我们的研究计划旨在确定控制 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 cki-1（细胞周期蛋白依赖性激酶抑制剂 p27Kip1 家族的成员）一起在调节细胞周期静止的网络中发挥作用。我们的策略应用线虫出色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，以确定将发育信号与 G1 进程控制联系起来的途径：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指定暂时的细胞周期静止。我们重点关注指导 cdc-14 活性细胞类型规范的两个过程：限制表达和亚细胞区室化。这些分析利用了一系列经过修饰的 cdc-14 转基因，旨在操纵 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) 在反向遗传筛选中鉴定在 cdc-14 介导的网络中起作用的其他基因，以发现控制 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缺陷，从而实现高效、简单的识别。 3) 使用在所述筛选中鉴定的基因开发遗传层次。该网络的组织将基于定义遗传相互作用、对 CDC-14 表达的影响以及新基因的亚细胞定位的测试。总之，拟议的研究将确定发育过程中控制 cdc-14 活性的途径，并将提供对用于将细胞周期调节与生长和发育相结合的一般机制的见解。该提案的目标是了解用于控制 cdc-14 活性的机制，cdc-14 是一种在暂时阻止细胞分裂方面发挥重要作用的基因。这些结果可能有助于了解肿瘤是如何形成的，因为癌症的特点是控制细胞分裂的基因缺陷。