G Protein Regulation of Golgi Structure and Function

G 蛋白对高尔基体结构和功能的调节

基本信息

批准号：
9926920
负责人：
PHILIP B WEDEGAERTNER
金额：
$ 31.2万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-05-07 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9926920
关键词：
Address Binding Proteins Biochemical Biological Assay Biosensor Blindness Cardiovascular Physiology Cell Cycle Checkpoint Cell Differentiation process Cell membrane Cell physiology Cells Couples Cultured Cells Disease Disease Pathway Endocrine System Diseases Fluorescence Microscopy G-Protein Signaling Pathway G-Protein-Coupled Receptors GTP-Binding Proteins Generations Golgi Apparatus Heart Diseases Heterotrimeric GTP-Binding Proteins Hypertension Immune Immunofluorescence Microscopy Infection Knowledge Link Location Malignant Neoplasms Mediating Mediator of activation protein Membrane Microtubules Mitosis Mitotic Molecular Morphology Mutation Analysis Neoplasm Metastasis Neurodegenerative Disorders Neurotransmitters Nocodazole Organelles Pathway interactions Pharmacology Physiological Physiological Processes Plasma Cells Play Process Proteins Regulation Research Research Proposals Role Signal Pathway Signal Transduction Signaling Protein Site Small Interfering RNA Smell Perception Structure Superoxide Dismutase Surface Taste Perception Testing Textbooks Vision Work cell motility experimental study extracellular inhibitor/antagonist live cell microscopy mutant nervous system disorder neuron development novel protein function protein protein interaction recruit response stathmin therapeutic target trafficking vesicle transport

项目摘要

Abstract Heterotrimeric G proteins () are well known for their function in linking G protein-coupled receptors (GPCRs) to a variety of intracellular responses, and thereby playing essential roles in transmitting a wide variety of extracellular signals into regulation of countless physiological processes. In the textbook view, G proteins carry out their function while associated with the cytoplasmic surface of a cell’s plasma membrane. In contrast to the classical view of plasma membrane-limited G protein signaling, it is becoming increasingly recognized that G protein localization is dynamic and regulated, such that they can reversibly traffic from the plasma membrane to intracellular locations, and that G proteins can have important cellular functions at intracellular sites. The research in this proposal focuses on understanding non-canonical functions of G subunits, and specifically roles for G in regulating signaling at the Golgi. Our previous work revealed an important role for Golgi-localized G in regulating a signaling pathway on the cytoplasmic surface of Golgi membranes that controls the Golgi exit of select protein cargo destined for the plasma membrane. The research in this current proposal will examine the hypothesis that G regulates signaling pathways that control Golgi integrity by regulating the fragmentation of the Golgi under physiological and pathophysiological conditions. Reversible Golgi fragmentation is a cellular phenomenon that occurs under normal conditions, such as during mitosis, and that occurs in disease states, such as infection, cancer and neurodegenerative disease. This application will focus on defining a novel role for G in regulating the mitotic Golgi fragmentation checkpoint and by interrogating a novel role for G in regulating microtubule- dependent Golgi fragmentation. In addition, this application will define upstream mechanisms that directly promote signaling by G at the Golgi. These objectives will be pursued by a variety of experimental approaches, including cultured cells, immunofluorescence microscopy, fluorescence microscopy of live cells, biosensors, pharmacological inhibitors, mutational analysis, and biochemical assays.

抽象的异三聚体 G 蛋白 () 以其连接 G 蛋白偶联受体的功能而闻名（GPCR）对多种细胞内反应，从而在广泛的传播中发挥重要作用在教科书看来，G将各种细胞外信号转化为无数生理过程的调节。蛋白质在与细胞质膜的细胞质表面结合时发挥其功能。与质膜限制 G 蛋白信号传导的经典观点相反，它正变得越来越重要。认识到 G 蛋白定位是动态且受调节的，因此它们可以可逆地从质膜到细胞内位置，并且 G 蛋白可以在以下位置具有重要的细胞功能：本提案的研究重点是了解 G 的非规范功能。亚基，以及 G 在调节高尔基体信号传导中的具体作用。高尔基体定位的 G 在调节高尔基体细胞质表面信号通路中的重要作用控制前往质膜的选定蛋白质货物的高尔基体出口的膜。当前提案中的研究将检验 G 调节信号通路的假设通过调节生理和条件下高尔基体的破碎来控制高尔基体的完整性可逆的高尔基体断裂是一种发生在病理生理条件下的细胞现象。正常情况下，例如有丝分裂期间，以及疾病状态下发生的情况，例如感染、癌症和该应用将重点定义 G 在调节神经退行性疾病方面的新作用。有丝分裂高尔基体碎片检查点并探讨 G 在调节微管中的新作用此外，该应用程序将定义直接的上游机制。促进高尔基体 G 的信号传导这些目标将通过各种实验来实现。方法，包括培养细胞、免疫荧光显微镜、活体荧光显微镜细胞、生物传感器、药理学抑制剂、突变分析和生化测定。