Regulation of Rho Family GTPases by G Proteins

G 蛋白对 Rho 家族 GTP 酶的调节

• 批准号: 7263215
• 负责人: TOHRU KOZASA
• 金额: $ 27.31万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7263215
• 关键词: Actins; Affect; Amino Acids; Antibodies; Appendix; Baculoviruses; Binding; Binding Proteins; Biochemical; Biological Assay; Caenorhabditis elegans; Cell Line; Cell physiology; Cells; Chimera organism; Chimeric Proteins; Complex; Couples; Critiques; Cytoskeleton; Data; Defect; Dominant-Negative Mutation; Embryonic Development; Family; Figs - dietary; GTP-Binding Protein Regulators; GTP-Binding Proteins; GTPase-Activating Proteins; Gene Expression; Generations; Goals; Green Fluorescent Proteins; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Hand; Hela Cells; Heterotrimeric GTP-Binding Proteins; Immunoblotting; Ligands; Mammalian Cell; Maps; Mediating; Membrane; Methods; Monitor; Neurites; Neurons; PC12 Cells; PDZ protein; PH Domain; Pathway interactions; Phenotype; Physiological; Process; Protein Family; Protein Overexpression; Protein Tyrosine Kinase; Proteins; Published Comment; RGS Domain; RGS Proteins; Range; Recombinant Proteins; Recombinants; Regulation; Resolution; Role; Signal Pathway; Signal Transduction; Source; Structure; System; Thrombin; Trypsin; Tyrosine; Tyrosine Phosphorylation; Tyrosine Phosphorylation Site; Work; X-Ray Crystallography; base; cell transformation; lysophosphatidic acid; member; mutant; novel; receptor; receptor binding; reconstitution; research study; response; rho; rho guanine nucleotide exchange factor p115; Actins; Affect; Amino Acids; Antibodies; Appendix; Baculoviruses; Binding; Binding Proteins; Biochemical; Biological Assay; Caenorhabditis elegans; Cell Line; Cell physiology; Cells; Chimera organism; Chimeric Proteins; Complex; Couples; Critiques; Cytoskeleton; Data; Defect; Dominant-Negative Mutation; Embryonic Development; Family; Figs - dietary; GTP-Binding Protein Regulators; GTP-Binding Proteins; GTPase-Activating Proteins; Gene Expression; Generations; Goals; Green Fluorescent Proteins; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate Phosphohydrolases; Hand; Hela Cells; Heterotrimeric GTP-Binding Proteins; Immunoblotting; Ligands; Mammalian Cell; Maps; Mediating; Membrane; Methods; Monitor; Neurites; Neurons; PC12 Cells; PDZ protein; PH Domain; Pathway interactions; Phenotype; Physiological; Process; Protein Family; Protein Overexpression; Protein Tyrosine Kinase; Proteins; Published Comment; RGS Domain; RGS Proteins; Range; Recombinant Proteins; Recombinants; Regulation; Resolution; Role; Signal Pathway; Signal Transduction; Source; Structure; System; Thrombin; Trypsin; Tyrosine; Tyrosine Phosphorylation; Tyrosine Phosphorylation Site; Work; X-Ray Crystallography; base; cell transformation; lysophosphatidic acid; member; mutant; novel; receptor; receptor binding; reconstitution; research study; response; rho; rho guanine nucleotide exchange factor p115

DESCRIPTION (provided by applicant): Heterotrimeric G proteins transduce a variety of signals from heptahetical membrane-bound receptors to intracellular effectors. Members of RGS (Regulator of G protein signaling) protein family regulate G protein mediated signals. Members of the Rho family of monomeric GTPases control the organization of the actin cytoskeleton and are involved in a variety of cellular functions. The objective of this proposal is to understand the mechanism of regulation of Rho family GTPases through heterotrimeric G protein-mediated signaling pathways. Among heterotrimeric G proteins, Galpha12 and Galpha13 participate in cellular processes such as cell transformation, neurite retraction, or embryonic development. Rho activation is involved in these signaling pathways. The immediate goal of this proposal is to understand biochemical mechanisms of regulation of the activity of Rho by Galpha12 and Galpha13. Rho family GTPases are regulated by guanine nucleotide exchange factors (GEFs). RhoGEFs with amino terminal RGS domain (RGS-RhoGEFs), p115RhoGEF, LARG, and PDZ-RhoGEF, were identified to function as effectors for Galpha12 and Galpha13. Galpha12/13 interacts with RGS domain of these RhoGEFs and regulates their RhoGEF activity. RGS domains of p115 and LARG act as GTPase activating proteins specific for Galpha12 and Galpha13. The structure basis of the mechanism of regulation of RGS-RhoGEFs by Galpha12 and Galpha13 will be analyzed. The crystal structures of Galpha12 or Galpha13 and its complex with LARG will be solved. The regions of Galpha12/13 or LARG that are involved in their functional interaction will be characterized. The biochemical mechanisms of regulation of RhoGEF activity of LARG or PDZ-RhoGEF will be investigated. Furthermore, Galpha12/13-LARG/PDZRhoGEF signaling pathway in neuronal cells will be analyzed. Galpha12-RhoGEF pathway was identified in C. elegans and its involvement in neuronal function and embryonic development was demonstrated. The physiological significance of this pathway in C. elegans will be investigated using genetical and biochemical methods.

描述（由申请人提供）：异三聚体G蛋白将各种信号从七个膜结合的受体转移到细胞内效应子。 RGS的成员（G蛋白信号传导的调节剂）蛋白家族调节G蛋白介导的信号。单体GTPases的Rho家族的成员控制肌动蛋白细胞骨架的组织，并参与多种细胞功能。该建议的目的是了解通过异三聚体G蛋白介导的信号通路调节Rho家族GTPase的机理。在异源三聚合物G蛋白中，Galpha12和Galpha13参与细胞过程，例如细胞转化，神经突收缩或胚胎发育。这些信号通路涉及RHO激活。该提案的直接目标是了解Galpha12和Galpha13对Rho活性调节的生化机制。 RHO家族GTPase受鸟嘌呤核苷酸交换因子（GEF）的调节。鉴定出具有氨基末端RGS域（RGS-RHOGEFS），P115RHOGEF，LARG和PDZ-RHOGEF的Rhogefs可作为Galpha12和Galpha13的效应子。 Galpha12/13与这些RhoGEF的RGS结构域相互作用，并调节其Rhogef活性。 P115和LARG的RGS结构域作为GTPase激活了对Galpha12和Galpha13的蛋白质。将分析Galpha12和Galpha13调节RGS-RHOGEF机制的结构基础。 Galpha12或Galpha13的晶体结构及其与LARG的复合物将得到解决。将表征与其功能相互作用有关的Galpha12/13或LARG的区域。将研究LARG或PDZ-RHOGEF的RhoGEF活性调节的生化机制。此外，将分析神经元细胞中的galpha12/13-larg/pdzrhogef信号通路。在秀丽隐杆线虫中鉴定出Galpha12-Rhogef途径，并证明了其参与神经元功能和胚胎发育。该途径在秀丽隐杆线虫中的生理意义将使用遗传和生化方法研究。