Structure and Function of the Exocyst Complex

外囊复合体的结构和功能

• 批准号: 7647934
• 负责人: Mary Munson
• 金额: $ 29.28万
• 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 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7647934
• 关键词: Address; Alleles; Animal Model; Binding; Biochemical; Biological; Biological Assay; Biological Models; C-terminal; Cell Cycle Proteins; Cell Survival; Cell membrane; Cells; Chimeric Proteins; Complex; Data; Data Set; Defect; Dimerization; Eukaryota; Eukaryotic Cell; Event; Exocytosis; Genes; Genetic; Genetic Screening; Goals; Growth; Hormones; Human; In Vitro; Knowledge; Location; Maintenance; Maps; Membrane; Membrane Fusion; Methods; Molecular; Movement; Mutation; Neurons; Organelles; Point Mutation; Process; Property; Proteins; Regulation; Research; Research Personnel; Resolution; Role; SNAP receptor; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins; Secretory Vesicles; Site; Specificity; Structure; Techniques; Testing; Transport Vesicles; Variant; Vesicle; Yeasts; base; cell growth; design; dimer; genetic regulatory protein; in vivo; interdisciplinary approach; man; mutant; neurotransmission; novel; programs; research study; rho GTP-Binding Proteins; target SNARE proteins; temperature sensitive mutant; trafficking

DESCRIPTION (provided by applicant): Eukaryotic cells utilize small membrane-bound vesicles to transport cargo between subcellular organelles, and to the plasma membrane for secretion. The proper function and specificity of the vesicular transport and membrane fusion processes are crucial for maintenance of cellular integrity, growth, cellular movement and secretory events such as hormone release and neurotransmission. Vesicle transport and fusion at the plasma membrane require many essential proteins, including the SNARE proteins and Sec1p that are involved in the membrane fusion process, the Rab and Rho GTPases, and a large complex called the exocyst. The exocyst complex has been implicated in a number of different functions: selection of the site of exocytosis; physical tethering of secretory vesicles to these sites; communicating with cytoskeletal and cell cycle proteins; and regulating the specificity and assembly of the SNARE proteins. None of these are well understood at the molecular level. Our aim is to combine biochemical and biophysical techniques with genetics and cell biological methods in order to understand the molecular functions of the exocyst complex. We have chosen to study the exocyst proteins from the model organism Saccharomyces cerevisiae so as to take advantage of the wealth of genetic and cell biological techniques available. To accomplish this goal, we have purified the exocyst protein Sec6p and are i) Investigating the structure of Sec6p by biochemical, biophysical and x-ray crystallographic studies; ii) Analyzing the function of Sec6p in vivo through characterization of specific Sec6p mutants; and (iii) Testing the effect of Sec6p on SNARE complex assembly. 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（由申请人提供）：真核细胞利用膜结合的小囊泡在亚细胞器之间运输货物，并在质膜之间进行分泌。囊泡转运和膜融合过程的适当功能和特异性对于维持细胞完整性，生长，细胞运动和分泌事件（例如激素释放和神经传递）至关重要。质膜处的囊泡转运和融合需要许多必需蛋白，包括膜融合过程中涉及的SNARE蛋白和SEC1P，Rab和Rho GTPases，以及一种称为外旋蛋白的大复合物。外囊复合物与许多不同的功能有关：胞吞位点的选择；将分泌囊泡的物理绑定到这些位点；与细胞骨架和细胞周期蛋白进行通信；并调节军鼓蛋白的特异性和组装。在分子水平上，这些都没有得到充分理解。我们的目的是将生化和生物物理技术与遗传学和细胞生物学方法相结合，以了解外囊复合物的分子功能。我们选择研究了酿酒酵母模型的催化蛋白，以利用可用的遗传和细胞生物学技术。为了实现这一目标，我们已经纯化了外囊蛋白SEC6P，并且是i）通过生化，生物物理和X射线晶体学研究研究SEC6P的结构； ii）通过表征特定的SEC6P突变体来分析SEC6P在体内的功能； （iii）测试SEC6P对SNARE复合物组件的影响。由于这些蛋白质是从酵母到人神经元保守的，因此这项研究将促进我们对所有真核细胞中分泌和生长如何受到调节的了解。