Mechanism of centrosome regulation from the Golgi

高尔基体调节中心体的机制

• 批准号: 8462130
• 负责人: CHRISTINE SUETTERLIN
• 金额: $ 27万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8462130
• 关键词: Actins; Affinity; Binding; Binding Proteins; Biological Assay; Cell physiology; Cells; Centrosome; Complex; Defect; Dominant-Negative Mutation; Dynein ATPase; Event; Fluorescence; Genetic; Golgi Apparatus; Guanine Nucleotide Exchange Factors; Human; Immunofluorescence Immunologic; In Vitro; Laboratories; Lead; Link; Location; Malignant Neoplasms; Mammalian Cell; Measures; Mediating; Monomeric GTP-Binding Proteins; Motor; Neoplasm Metastasis; Pathway interactions; Peripheral; Phenotype; Photons; Plants; Process; Proteins; Recruitment Activity; Regulation; Regulatory Pathway; Signal Transduction; Spectrum Analysis; Testing; Tissue Differentiation; Tissues; Work; Yeasts; cell motility; cell transformation; dynactin; fly; in vivo; novel; overexpression; protein complex; public health relevance; rho GTP-Binding Proteins

DESCRIPTION (provided by applicant): The objective of this project is to understand a control mechanism that regulates the centrosome from the adjacent Golgi apparatus in mammalian cells. While the proximity of the Golgi and centrosome is unique to mammalian cells, and not observed in yeast, plant or fly cells, its functional significance has been elusive. Work in our laboratory has identified a centrosome regulatory pathway in which a Golgi protein, GM130, causes a guanine nucleotide exchange factor called Tuba to activate Cdc42 at the Golgi. This project seeks to determine how these events at the Golgi control centrosome organization and function and whether they depend on Golgi-centrosome proximity. Aim 1 will determine whether GM130 activates Cdc42 at the Golgi by increasing the binding affinity of Tuba for Cdc42 or by recruiting Cdc42 to the Golgi. This aim will also examine if GM130 is the major regulator of Cdc42 at the Golgi by assaying whether GM130 controls additional cellular processes at the Golgi that are known to be regulated by Cdc42, such as Golgi to ER transport and local actin assembly at the Golgi. A potential negative regulator of Golgi-associated Cdc42 will also be studied. Aim 2 will examine how the GM130-Cdc42 pathway exerts its effect on the centrosome. A Cdc42 effector, Par6?, will be studied to determine if it is a downstream component of this regulatory pathway and if it is transported to the centrosome by the motor protein, dynein, through interactions with the dynactin subunit, p150Glued. PCM-1, a protein that recruits additional proteins to the centrosome, will be studied to determine if it acts downstream of Par6? to mediate the control of centrosome organization by the GM130-Cdc42 pathway. Aim 3 will study the functional significance of Golgi-centrosome proximity by examining if Cdc42 can be mobilized from the Golgi to the centrosome so that it can regulate centrosome organization. The pericentriolar location of the Golgi will be disrupted as another means of testing whether Golgi centrosome proximity is necessary for regulation of centrosome organization by the GM130-Cdc42 pathway.

描述（由申请人提供）：该项目的目的是了解一种控制机制，该控制机制可调节哺乳动物细胞中相邻高尔基体的中心体。虽然高尔基体和中心体的接近性是哺乳动物细胞独有的，并且在酵母，植物或飞蝇细胞中没有观察到，但其功能意义是难以捉摸的。我们实验室的工作已经确定了一种中心体调节途径，其中高尔基蛋白GM130导致称为Tuba的鸟嘌呤核苷酸交换因子在高尔基体上激活Cdc42。该项目旨在确定高尔基控制中心体组织和功能中的这些事件如何依赖于高尔基体中心体的接近度。 AIM 1将通过增加TUBA对Cdc42的结合亲和力还是通过将Cdc42募集到高尔基体来确定GM130是否会激活高尔基体的Cdc42。该目标还将检查GM130是否是GOLGI中CDC42的主要调节剂，通过测定GM130是否控制GOLGI的其他细胞过程，该过程已知受CDC42的调节，例如Golgi到Golgi的ER运输和局部肌动蛋白组装。还将研究高尔基相关CDC42的潜在负调节剂。 AIM 2将检查GM130-CDC42途径如何对中心体施加影响。将研究Cdc42效应子，PAR6？，以确定它是否是该调节途径的下游组成部分，以及是否通过与Dynactin subunit的相互作用P150Glued通过运动蛋白Dynein转运到中心体。 PCM-1是一种将其他蛋白质募集到中心体的蛋白质，将研究以确定它是否在PAR6下游起作用？通过GM130-CDC42途径调解中心体组织的控制。 AIM 3将通过检查Cdc42是否可以从高尔基体动员到中心体，以便它可以调节中心体组织。高尔基人的周围三醇位置将被破坏，作为测试高尔基体邻近近端是否需要通过GM130-CDC42途径调节中心体组织的另一种手段。