A transmembrane Rab GTPase accelerating protein targeted to peroxisomes

一种针对过氧化物酶体的跨膜 Rab GTP 酶加速蛋白

• 批准号: 10398972
• 负责人: Daniel P. Nickerson
• 金额: $ 14.7万
• 依托单位: CALIFORNIA STATE UNIV SAN BERNARDINO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10398972
• 关键词: 3-Dimensional; Address; Affinity; Alleles; Area; Autophagocytosis; Biochemical; Biological Assay; Cardiovascular Diseases; Cardiovascular system; Cells; Cellular biology; Client; Coupled; Critical Pathways; Development; Disease; Docking; Endoplasmic Reticulum; Enzymes; Eukaryotic Cell; Exercise; Family; Fluorescence Microscopy; Foundations; GTP Binding; Genes; Goals; Gold; Golgi Apparatus; Growth; Guanosine Triphosphate Phosphohydrolases; Health; Human; Imaging Techniques; Integral Membrane Protein; Kinetics; Lipids; Maintenance; Maps; Mass Spectrum Analysis; Membrane; Membrane Fusion; Metabolic Diseases; Metabolic Pathway; Metabolism; Microscopic; Mitochondria; Mitochondrial Proteins; Molecular; Molecular Chaperones; Morphology; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Organelles; Organism; Orthologous Gene; Outcomes Research; Pathway interactions; Phenotype; Play; Process; Proteins; Proteomics; Quality Control; Regulation; Reporting; Research; Resources; Role; Route; Saccharomyces cerevisiae; Signal Transduction; Signaling Protein; Source; Stimulus; Stress; Synthetic Genes; Systems Biology; Technology; Testing; Therapeutic Intervention; Time; Toxic effect; Transmembrane Transport; Vesicle; Yeasts; age related; biological adaptation to stress; electron tomography; in vivo; innovation; lipid metabolism; membrane biogenesis; peroxisome; prevent; programs; rab GTP-Binding Proteins; repaired; response; spatiotemporal; targeted treatment; vesicle transport; yeast genome

Project Summary Understanding how cells adapt to stress and repair damage is one of the highest priorities in cell biology and health research. The endoplasmic reticulum (ER) is a key source of lipid and proteins for building and maintaining several organelles in cells, and in response to stimuli it is capable of directing resources into an assortment of transport pathways. The signaling protein Ypt1/Rab1 is a Rab GTPase (Rab) that plays essential roles in how the ER directs resources and executes quality control of damaged organelles including mitochondria and peroxisomes. Our long-term goal is to understand the mechanisms for how lipid and protein cargos are routed and re-routed to specific itineraries in response to cellular stimuli and stresses, which will inform development of targeted therapeutic interventions to address diseases. We have found that the GTPase accelerating protein Gyp8, an evolutionarily conserved but poorly understood negative regulator of Ypt1/Rab1 signaling, localizes to the ER, peroxisomes and mitochondria and impinges on Ypt1 signaling. Our central hypotheses are that Gyp8 functions to modulate Ypt1/Rab1 signaling in the early secretory pathway (ER) and at non-secretory membranes (peroxisomes and mitochondria) that are subject to frequent damage and quality control via Ypt1/Rab1-dependent selective autophagy. Also, since Gyp8 localizes to organelles that commonly tether/dock to exchange materials in essential metabolic pathways, we predict that Gyp8 regulates Rab-dependent tethering interactions, particularly among organelles specialized for lipid storage and metabolism. To test our central hypotheses and advance understanding of several membrane biogenesis pathways that originate at the ER, we will pursue these specific aims: 1) Identify intra- and extra-genic factors that control the subcellular itinerary and activity of Gyp8; 2) Determine target Rab GTPase(s) regulated by Gyp8 in the secretory pathway; and 3) Define the role of Gyp8 in regulating peroxisomal and mitochondrial dynamics. The proposed research is innovative both for area of focus and technical approach. While Ypt1/Rab1 signaling controls multiple transport pathways essential to cellular health, understanding of where and when signal must be terminated to accomplish each of its roles is particularly incomplete. The experimental plan combines gold standard biochemical and imaging techniques in organelle and vesicular transport (enzyme-coupled kinetic transport assays and 3D electron tomography) with systems biology approaches (synthetic gene array and affinity capture mass spectrometry proteomics). The proposed research is significant because defining how Ypt1/Rab1 is regulated to support and exercise quality control of ER, mitochondria and peroxisomes is foundational to understanding aspects of cardiovascular, neurodegenerative and metabolic disorders.

项目概要 了解细胞如何适应压力和修复损伤是细胞生物学的最高优先事项之一 健康研究。内质网 (ER) 是脂质和蛋白质的关键来源，用于构建和 维持细胞中的多种细胞器，并且响应刺激，它能够将资源引导到 运输路径的分类。信号蛋白 Ypt1/Rab1 是一种 Rab GTPase (Rab)，其作用是 急诊室在如何引导资源和对受损细胞器进行质量控制方面发挥着重要作用，包括 线粒体和过氧化物酶体。我们的长期目标是了解脂质和蛋白质如何发挥作用的机制 货物根据细胞刺激和压力被路由和重新路由到特定的行程，这将 为制定针对疾病的有针对性的治疗干预措施提供信息。我们发现 GTPase 加速蛋白 Gyp8，一种进化上保守但知之甚少的负调节因子 Ypt1/Rab1 信号传导定位于 ER、过氧化物酶体和线粒体，并影响 Ypt1 信号传导。我们的 中心假设是 Gyp8 在早期分泌途径中调节 Ypt1/Rab1 信号传导 (ER) 和经常受到损伤的非分泌膜（过氧化物酶体和线粒体） 通过 Ypt1/Rab1 依赖性选择性自噬进行质量控制。此外，由于 Gyp8 定位于细胞器 通常在重要的代谢途径中连接/对接以交换物质，我们预测 Gyp8 调节 Rab 依赖性束缚相互作用，特别是专门用于脂质储存和 代谢。测试我们的中心假设并加深对几种膜生物发生的理解 起源于 ER 的途径，我们将追求以下具体目标：1）识别基因内和基因外因素 控制 Gyp8 的亚细胞行程和活性； 2) 确定受以下因素调节的目标 Rab GTPase Gyp8 在分泌途径中； 3) 定义 Gyp8 在调节过氧化物酶体和线粒体中的作用 动力学。拟议的研究在重点领域和技术方法上均具有创新性。尽管 Ypt1/Rab1 信号传导控制对细胞健康至关重要的多种运输途径，了解在哪里 当必须终止信号才能完成其每个角色时，这是特别不完整的。这 实验计划结合了细胞器和囊泡的金标准生化和成像技术 运输（酶联动运输分析和 3D 电子断层扫描）与系统生物学 方法（合成基因阵列和亲和捕获质谱蛋白质组学）。拟议的研究 很重要，因为定义如何调节 Ypt1/Rab1 以支持和实施 ER 质量控制， 线粒体和过氧化物酶体是理解心血管、神经退行性疾病方面的基础 和代谢紊乱。