Biochemical regulation of the ras-like protein, CDC42

ras 样蛋白 CDC42 的生化调控

• 批准号: 7221221
• 负责人: RICHARD A. CERIONE
• 金额: $ 28.46万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7221221
• 关键词: Actins; Address; Area; B-Cell Lymphomas; Binding; Binding Proteins; Biochemical; Biochemical Genetics; Biological Assay; CDC42 gene; Cell Cycle Progression; Cell Polarity; Cell membrane; Cell physiology; Cells; Cellular biology; Clathrin; Coat Protein Complex I; Coatomer gamma Subunit; Complex; Cytokinesis; Cytoskeleton; Diffuse; Dissociation; Docking; Endoplasmic Reticulum; Epidermal Growth Factor Receptor; Event; Family; Funding; GDP dissociation inhibitor; GTP Binding; GTP-Binding Proteins; Genetic; Glycoproteins; Goals; Golgi Apparatus; Growth Factor Receptors; Guanine Nucleotide Dissociation Inhibitors; Guanine Nucleotide Exchange Factors; Guanosine Diphosphate; Guanosine Triphosphate Phosphohydrolases; Homologous Gene; Human; Laboratories; Lead; Learning; Libraries; Link; Localized; Location; Mammalian Cell; Mediating; Membrane; Methods; Modeling; Molecular; Movement; Mutation; Nude Mice; Numbers; Phosphotransferases; Play; Principal Investigator; Process; Protein Family; Proteins; Range; Receptor Down-Regulation; Recycling; Regulation; Resolution; Roentgen Rays; Role; Running; Saccharomyces cerevisiae; Series; Signal Pathway; Signal Transduction; Signaling Protein; Site; Small Interfering RNA; Sorting - Cell Movement; Stomatitis; Testing; Vesicle; Viral; Work; Yeasts; base; cdc42 GTP-Binding Protein; cell growth; cell growth regulation; cell motility; cellular targeting; design; genetic regulatory protein; inhibitor/antagonist; insight; interest; intersectin 1; member; metaplastic cell transformation; mouse Gdi2 protein; mutant; nexin; programs; ras-Related G-Proteins; response; rho; structural biology; three dimensional structure; trafficking; tumor

DESCRIPTION (provided by applicant): The Ras-related GTP-binding protein Cdc42 has been implicated in a number of fundamentally important cellular processes including the establishment of cell polarity and motility through influences on the actin cytoskeleton, and the regulation of cell-cycle progression. The tight regulation of the GTP-binding/GTPase cycle of Cdc42 is essential to its cellular functions. Mutations that trap Cdc42 in either the GDP-bound or GTP-bound state inhibit cell growth, whereas those that give rise to an accelerated cycling of Cdc42 between these states lead to cellular transformation and tumor formation in nude mice. During the past funding period, we have combined biochemical, genetic, and structural biology-based approaches to study the regulation of Cdc42 by guanine nucleotide exchange factors and the Rho-GDP-dissociation inhibitor (RhoGDI), as well as examined the interactions of activated Cdc42 with various downstream signaling targets, including ACK (Activated Cdc42- associated kinase) and IQGAP. In addition, we have identified two new targets for Cdc42, the gamma-coatomer subunit (gammaCOP) of the COPI complex and the p85Cool-1 (for Cloned-out of library)/beta-Pix (PAK-interactive exchange factor) protein. These studies have provided us with new information regarding the molecular mechanisms underlying the cellular regulation and function of Cdc42. However, perhaps most important, work during the past funding period has raised an interesting and somewhat unanticipated role for Cdc42, namely linking intracellular trafficking activities to cell signaling and cell growth regulation. In this renewal application, we plan to examine this interesting new role for Cdc42 by focusing on those upstream regulators and downstream target/effectors that appear to implicate most strongly Cdc42 in cellular trafficking functions. This will constitute 4 lines of study. Aim la.) Identify and characterize the protein complexes that couple the activation of Cdc42 to the sorting and trafficking of EGF receptors. Aim lb.) Determine why Cdc42-gammaCOP interactions are essential for Cdc42-mediated cellular transformation. Aim 2a.) Determine whether IQGAP serves as a docking site for Cdc42 and other proteins involved in intracellular trafficking functions. Aim 2b.) Determine whether RhoGDI plays a fundamental role in Cdc42-mediated intracellular trafficking activities. These studies are expected to yield important new insights into a critical cellular function for Cdc42 that may form the basis for a diversity of Cdc42-mediated cellular responses ranging from cell cycle progression and cellular transformation to morphological and actin cytoskeletal changes.

描述（由申请人提供）：与RAS相关的GTP结合蛋白CDC42与许多根本重要的细胞过程有关，包括通过对肌动蛋白细胞骨架的影响以及细胞周期进展的调节，包括建立细胞极性和运动性。 Cdc42的GTP结合/GTPase周期的严格调节对于其细胞功能至关重要。在GDP结合或GTP结合的状态中捕获CDC42的突变抑制细胞的生长，而这些态在这些状态之间会加速Cdc42加速的细胞生长，从而导致细胞转化和裸体小鼠的肿瘤形成。在过去的资金期间，我们结合了基于生物化学，遗传和结构生物学的方法来研究鸟嘌呤核苷酸交换因子对CDC42的调节，而Rho-GDP分解抑制剂（RHOGDI）（RHOGDI），并检查了激活的CDC42与各种cDC的相互作用，包括Aciptrems Signalsing cdc42（Activects a Activect ancipts ccin）。此外，我们已经确定了CDC42的两个新靶标，COPI复合物的伽马型亚基（Gammacop）和P85Cool-1（用于克隆的文库）/β-Pix（PAK-Interactive Exchange frossive因子）蛋白质。这些研究为我们提供了有关CDC42细胞调节和功能的分子机制的新信息。但是，也许最重要的是，在过去的资金期间的工作提高了CDC42的有趣且意外的作用，即将细胞内运输活动与细胞信号传导和细胞生长调节联系起来。在此续订应用中，我们计划通过关注那些上游调节器和下游目标/效应子来研究CDC42的这种有趣的新作用，这些目标和下游目标/效应子似乎在细胞运输函数中牵涉到最强烈的CDC42。这将构成4条研究。 AIM LA。）确定并表征将Cdc42激活与EGF受体的分类和运输相结合的蛋白质复合物。 AIM磅）确定为什么CDC42-Gammacop相互作用对于CDC42介导的细胞转化至关重要。 AIM 2A。）确定IQGAP是否充当CDC42和其他参与细胞内运输功能的蛋白质的对接位点。 AIM 2B。）确定Rhogdi是否在CDC42介导的细胞内贩运活动中起着基本作用。预计这些研究将对CDC42的关键细胞功能产生重要的新见解，这可能是CDC42介导的细胞反应的多样性的基础，从细胞周期进程和细胞转化到形态学和肌动蛋白细胞骨骼变化。