Structure and Function of the Exocyst Complex

外囊复合体的结构和功能

• 批准号: 8764547
• 负责人: Mary Munson
• 金额: $ 55.36万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8764547
• 关键词: Animal Model; Architecture; Binding; Biochemical; Biological; Biological Process; Cell membrane; Cells; Cellular Morphology; Cellular biology; Chimeric Proteins; Communication; Complement; Complex; Crystallography; Data; Development; Diabetes Mellitus; Dissection; Electron Microscopy; Eukaryota; Eukaryotic Cell; Event; Foundations; Funding; Genetic; Growth; Growth and Development function; Hormones; Human; Imagery; In Vitro; Individual; Investigation; Knowledge; Lead; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Membrane; Membrane Fusion; Methods; Modeling; Molecular; Molecular Models; Molecular Structure; Monomeric GTP-Binding Proteins; Movement; Negative Staining; Neurons; Organelles; Pathogenesis; Pathway interactions; Plant Roots; Process; Proteins; Publishing; Quality Control; Reagent; Regulation; Research; Resolution; Roentgen Rays; Role; SNAP receptor; Saccharomyces cerevisiae; Secretory Vesicles; Site; Specificity; Spectrum Analysis; Staging; Structure; Techniques; Technology; Testing; Time; Vesicle; Yeast Model System; Yeasts; biophysical techniques; cell growth; ciliopathy; crosslink; human disease; improved; in vivo; insight; man; molecular modeling; multidisciplinary; mutant; neurotransmission; novel; protein complex; public health relevance; rab GTP-Binding Proteins; rho GTP-Binding Proteins; single molecule; three dimensional structure; tool; trafficking; yeast genetics

DESCRIPTION (provided by applicant): 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 an octameric tethering complex called the exocyst. Although the exocyst complex has been implicated in a number of different functions involved in recognition, tethering and quality control of SNARE assembly and fusion, none of these are well understood at the molecular level. We are using a multidisciplinary strategy of biochemical and biophysical techniques, combined with genetics and cell biological methods, in order to understand the molecular architecture and function of the exocyst complex. 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. Our studies aim to: 1) map the functional organization of the exocyst complex through biochemical studies in vitro and analyze mutants to test the function of the exocyst in vivo; (2) determine the 3D structure of the entire exocyst complex using electron microscopy, crystallography and molecular modeling; and (3) watch the exocyst tether vesicles at the single molecule level to analyze the requirements for tethering, and (4) dissect the role of the exocyst and Sec1 in SNARE complex assembly and membrane fusion. 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.

描述（由申请方提供）：真核细胞使用小的膜结合囊泡作为载体，在亚细胞器之间运输货物，并将货物运输至质膜进行分泌。囊泡运输和膜融合过程的调节对于细胞形态、生长、运动和分泌，包括激素释放和神经传递至关重要。这些过程需要许多必需的蛋白质，包括参与膜融合过程的SNARE蛋白和Sec 1，Rab和Rho GTP酶，以及称为外囊的八聚体束缚复合物。尽管外囊复合物已经涉及到SNARE组装和融合的识别、拴系和质量控制中所涉及的许多不同功能，但这些功能在分子水平上都没有得到很好的理解。我们正在使用生物化学和生物物理技术的多学科策略，结合遗传学和细胞生物学方法，以了解外囊复合物的分子结构和功能。我们研究了模式生物酿酒酵母的外囊蛋白，以利用丰富的遗传，细胞生物学和生物化学技术。我们的研究旨在：1）通过体外生物化学研究绘制外囊复合物的功能组织，并分析突变体以测试外囊的体内功能;（2）使用电子显微镜、晶体学和分子建模确定整个外囊复合物的3D结构;和（3）在单分子水平上观察外囊系链囊泡以分析系链的要求，（4）分析外囊和Sec 1在SNARE复合物组装和膜融合中的作用。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中分泌和生长是如何调节的知识。