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: 10595549
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10595549
• 关键词: 3-Dimensional; Acceleration; Address; Affinity; Alleles; Area; Autophagocytosis; Binding; 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; GTP Binding; Genes; Goals; 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 Membrane Protein; Molecular; Molecular Chaperones; Morphology; Mutagens; 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; 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.

项目摘要 了解细胞如何适应压力和修复损伤是细胞生物学和 健康研究。内质网（ER）是建筑物和蛋白质的关键来源 保持细胞中的几个细胞器，并在响应刺激中，能够将资源引导到一个 各种运输途径。信号蛋白ypt1/rab1是播放的Rab GTPase（RAB） 急诊室如何指导资源和执行受损细胞器的质量控制在内的重要作用 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的机制 响应细胞刺激和应力，将货物路由并重新列为特定的行程，这将 告知有针对性的治疗干预措施以解决疾病。我们发现 GTPase加速蛋白GYP8，这是一种进化保守但知之甚少的负调节剂 YPT1/RAB1信号传导，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信号的功能 （ER）和在非分泌膜（过氧化物酶体和线粒体）处受到频繁损害 和质量控制通过YPT1/RAB1依赖性选择性自噬。另外，由于GYP8本地化为细胞器 通常是在基本代谢途径中交换材料的系绳/码头，我们预测GYP8 调节RAB依赖性的绑扎相互作用，尤其是在专门用于脂质存储和的细胞器中 代谢。测试我们的中心假设并提高对几种膜生物发生的理解 起源于急诊室的途径，我们将追求这些特定目的：1）确定内部和外部因素 控制GYP8的亚细胞行程和活性； 2）确定由 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。拟议的研究对于重点领域和技术方法都是创新的。尽管 YPT1/RAB1信号控制蜂窝健康必不可少的多个传输途径 当必须终止信号以完成其每个角色时，特别是不完整的。这 实验计划结合了细胞器和水泡中的金标准生化和成像技术 使用系统生物学的传输（酶耦合动力学传输测定和3D电子断层扫描） 方法（合成基因阵列和亲和力捕获质谱蛋白质组学）。拟议的研究 之所以重要，是因为定义如何调节YPT1/RAB1以支持和行使ER的质量控制， 线粒体和过氧化物酶体是理解心血管，神经退行性方面的基础 和代谢障碍。