Dissecting the Molecular Mechanisms of Exocytic Vesicle Tethering and Fusion

剖析胞吐囊泡束缚和融合的分子机制

• 批准号: 10552360
• 负责人: Mary Munson
• 金额: $ 66.91万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10552360
• 关键词: Address; Architecture; Binding; Biochemical; Biological; Biological Process; Cell membrane; Cells; Cellular Morphology; Communication; Complex; Development; Diabetes Mellitus; Disease; Eukaryotic Cell; Functional disorder; Genetic; Growth; Hormones; Human; Infection; Knowledge; Lipids; Malignant Neoplasms; Membrane; Membrane Fusion; Methods; Molecular; Movement; Neurons; Organelles; Organism; Process; Proteins; Quality Control; Reagent; Regulation; Research; Role; SNAP receptor; Saccharomyces cerevisiae; Specificity; Techniques; Technology; Vesicle; Yeasts; biophysical techniques; cell growth; ciliopathy; human disease; man; model organism; multidisciplinary; neurotransmission; pathogen; rab GTP-Binding Proteins; rho GTP-Binding Proteins; tool; vesicle transport

PROJECT SUMMARY Eukaryotic cells transport cargo between subcellular organelles, and to the plasma membrane for secretion, using small membrane-bound vesicles are carriers. The regulation of vesicular transport and membrane fusion processes are crucial for cellular morphology, growth, movement and secretion, including hormone release and neurotransmission. Many essential proteins are required for these processes, including the SNARE proteins and Sec1 that are involved in the membrane fusion process, the Rab and Rho GTPases, and the exocyst octameric tethering complex. Exocyst has been implicated in a number of different functions involved in recognition, tethering and quality control of SNARE assembly and fusion, but none of these are well understood at the molecular level. We use a multidisciplinary strategy of biochemical, structural and biophysical techniques, combined with genetics and cell biological methods, to understand the molecular architecture and function of the exocyst complex, Sec1 and regulation of SNARE complex assembly and fusion. We study the exocyst proteins from the model organism Saccharomyces cerevisiae to take advantage of the wealth of genetic, cell biological and biochemical techniques available, but have expanded to exocyst from other organisms. Our studies aim to address the following fundamental questions: How is the specificity of vesicle targeting and fusion achieved? How do exocyst and Sec1 function to regulate SNAREs? What are the roles for different partner proteins and lipids? What happens when exocyst and Sec1 functions are dysregulated? Because these proteins are conserved from yeast to human neurons, this research will advance our knowledge of how secretion and growth are regulated in all eukaryotic cells. We will also illuminate the molecular consequences of disease-associated dysfunction.

项目概要 真核细胞在亚细胞器之间运输货物，并运输到质膜进行分泌， 使用小的膜结合囊泡作为载体。囊泡运输和膜融合的调节 过程对于细胞形态、生长、运动和分泌（包括激素释放）至关重要 和神经传递。这些过程需要许多必需的蛋白质，包括 SNARE 参与膜融合过程的蛋白质和 Sec1、Rab 和 Rho GTPases 以及 外囊八聚体束缚复合物。 Exocyst 涉及许多不同的功能 SNARE 组装和融合的识别、束缚和质量控制，但这些都不是很好 在分子水平上理解。我们采用生物化学、结构和多学科策略 生物物理技术，结合遗传学和细胞生物学方法，了解分子 外囊复合体、Sec1 的结构和功能以及 SNARE 复合体组装和调节 融合。我们研究了模式生物酿酒酵母的外囊蛋白，以利用 现有丰富的遗传、细胞生物学和生化技术，但已扩展到外囊 来自其他生物体。我们的研究旨在解决以下基本问题： 囊泡靶向和融合实现了吗？ exocyst 和 Sec1 如何发挥作用来调节 SNARE？有哪些 不同伙伴蛋白和脂质的作用？当 exocyst 和 Sec1 功能发挥作用时会发生什么 失调？由于这些蛋白质从酵母到人类神经元都是保守的，因此这项研究将推进 我们对所有真核细胞如何调节分泌和生长的了解。我们还将照亮 疾病相关功能障碍的分子后果。