Structure and Function of the Exocyst Complex

外囊复合体的结构和功能

• 批准号: 10077638
• 负责人: Mary Munson
• 金额: $ 2.11万
• 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 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077638
• 关键词: Animal Model; Architecture; Auxins; Binding; Biochemical; Biological; Biological Process; Biophysics; Cell membrane; Cells; Cellular Morphology; Cellular biology; Chimeric Proteins; Communication; Complex; Cryoelectron Microscopy; Crystallography; Data; Development; Diabetes Mellitus; Dissection; Electron Microscopy; Eukaryota; Eukaryotic Cell; Event; Foundations; Funding; Genetic; Golgi Apparatus; Growth; Growth and Development function; Hormones; Human; In Vitro; Individual; Investigation; Knowledge; Lead; Liposomes; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Measures; Mechanics; Mediating; Membrane; Membrane Fusion; Methods; Modeling; Molecular; Molecular Conformation; Molecular Structure; Monomeric GTP-Binding Proteins; Movement; Myosin ATPase; Negative Staining; Neurons; Organelles; Organism; Pathogenesis; Pathway interactions; Plant Roots; Play; Process; Proteins; Publishing; Quality Control; Reagent; Recycling; Regulation; Research; Resolution; Role; SNAP receptor; Saccharomyces cerevisiae; Secretory Vesicles; Site; Specificity; Spectrum Analysis; Structure; System; Techniques; Technology; Testing; Time; Vesicle; Yeast Model System; Yeasts; biophysical techniques; cell growth; ciliopathy; crosslink; human disease; improved; in vitro Assay; in vivo; insight; man; molecular modeling; multidisciplinary; mutant; neurotransmission; novel; protein complex; rab GTP-Binding Proteins; rho GTP-Binding Proteins; single molecule; three dimensional structure; tool; trafficking; 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 a 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, but have expanded to exocyst from other organisms. 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.

项目摘要 真核细胞在亚细胞细胞器之间和质膜之间运输货物，以进行分泌， 使用小膜囊泡是载体。囊泡运输和膜融合的调节 过程对于细胞形态，生长，运动和分泌至关重要，包括激素释放 和神经传递。这些过程需要许多基本蛋白质，包括网罗 参与膜融合过程，RAB和RHO GTPases以及A的蛋白质和SEC1 八聚束缚复合物称为外囊。尽管外囊综合体已与 识别，绑扎和质量控制涉及的不同功能的数量，以及 融合，在分子水平上都没有很好地理解这些。我们正在使用一种多学科策略 生化和生物物理技术，结合遗传学和细胞生物学方法，以便为了 了解外囊复合物的分子结构和功能。我们研究外囊蛋白 从模型生物体酿酒酵母，以利用遗传，细胞生物学的丰富 以及可用的生化技术，但已从其他生物体扩展为外囊肿。我们的研究目的是： 1）通过体外的生化研究绘制胞外复合物的功能组织，并分析 突变体测试体内外囊肿的功能； （2）确定整个外囊复合体的3D结构 使用电子显微镜，晶体学和分子建模； （3）观看外囊的系绳囊泡 在单分子水平上分析束缚的要求，（4）剖析外囊肿的作用 和SENARE复合物组件和膜融合中的SEC1。因为这些蛋白质是从中保存的 对人类神经元的酵母，这项研究将提高我们对分泌和增长如何调节的了解 在所有真核细胞中。