G Protein Regulation of Golgi Structure and Function

G 蛋白对高尔基体结构和功能的调节

• 批准号: 10359763
• 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
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-07 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10359763
• 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; familial amyotrophic lateral sclerosis; inhibitor; live cell microscopy; mutant; nervous system disorder; neuron development; novel; protein function; protein protein interaction; recruit; response; stathmin; therapeutic target; trafficking; vesicle transport

Abstract Heterotrimeric G proteins (αβγ) 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 Gβγ subunits, and specifically roles for Gβγ in regulating signaling at the Golgi. Our previous work revealed an important role for Golgi-localized Gβγ 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 Gβγ 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 Gβγ in regulating the mitotic Golgi fragmentation checkpoint and by interrogating a novel role for Gβγ in regulating microtubule- dependent Golgi fragmentation. In addition, this application will define upstream mechanisms that directly promote signaling by Gβγ 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（αβγ）以连接G蛋白偶联受体的功能而闻名 （GPCR）到各种细胞内反应，从而在传播宽阔的地方发挥着重要作用 细胞外信号的多种多样，以调节无数的生理过程。在教科书视图中，G 蛋白质在与细胞质膜的细胞质表面相关的同时发挥其功能。 与质膜限制G蛋白信号传导的经典视图相反，它变得越来越多 认识到G蛋白定位是动态和调节的，因此它们可以可逆地从 质膜到细胞内的位置，G蛋白可以具有重要的细胞功能 细胞内部位。该提案中的研究重点是理解Gβγ的非规范功能 亚基，特别是Gβγ在高尔基体调节信号传导中的作用。我们以前的工作揭示了 高尔基体定位的Gβγ在高尔基体细胞质表面的信号通路中的重要作用 控制着针对质膜的精选蛋白质货物的高尔基体出口的膜。 当前建议中的研究将研究以下假设：Gβγ调节信号通路 通过在生理和 病理生理状况。可逆的高尔基碎片是一种细胞现象，发生在 正常情况，例如有丝分裂期间，并且发生在疾病状态，例如感染，癌症和 神经退行性疾病。该应用将着重于定义Gβγ在调节的新作用 有丝分裂高尔基分裂检查点，并通过审问Gβγ在调节微管中的新作用 依赖的高尔基分裂。此外，此应用程序将定义直接的上游机制 GOLGI促进Gβγ的信号传导。这些目标将由各种实验性追求 方法，包括培养的细胞，免疫荧光显微镜，生物的荧光显微镜 细胞，生物传感器，药物抑制剂，突变分析和生化测定。