Molecular Studies of Eukaryotic Cell Surface Growth

真核细胞表面生长的分子研究

• 批准号: 6769903
• 负责人: RANDY W. SCHEKMAN
• 金额: $ 17.77万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6769903
• 关键词: Golgi apparatus; SDS polyacrylamide gel electrophoresis; Saccharomyces cerevisiae; beta fructofuranosidase; cell cycle; cell membrane; chitin; clathrin; density gradient ultracentrifugation; developmental genetics; electron microscopy; enzyme activity; eukaryote; gene expression; genetic regulation; immunoelectron microscopy; immunoprecipitation; laboratory rabbit; membrane proteins; molecular cloning; protein purification; protein transport; secretory protein; vesicle /vacuole; western blottings

DESCRIPTION (provided by applicant): Protein secretion is a fundamentally conserved process that provides a mechanism for cells to create stable Intracellular compartments such as the ER, Golgi, lysosome and plasma membrane, and to communicate with other cells and tissues. A molecular mechanistic analysis of this process in Saccharomyces cerevisiae has uncovered genes and mechanisms that are shared among all eukaryotes. This proposal extends a new line of investigation that I initiated in my previous competitive renewal concerning the mechanism of vesicle traffic from the Golgi complex and particularly the pathway of traffic from the trans Golgi to the cell surface. Evidence has emerged to suggest multiple paths of vesicle formation from the trans Golgi, one of which involves sorting a subset of secretory and vacuolar proteins in the pre-vacuolar compartment, or late endosome. Another involves sorting of early endosomal resident proteins and biosynthetic forms of cell surface membrane proteins. Genetic and cell fractionation studies are proposed to discover the genes that govern sorting of cell surface precursors in the early and late endosomes. A genetic approach is described to identify the sorting signal on a membrane protein that is directed from the early endosome to the nascent division septum. A biochemical approach has been developed to explore the exact function of gene products associated with vesicle budding from the trans Golgi/early endosome in yeast. Transport vesicle formation has been reconstituted from lysates of yeast cells. Vesicle formation is assayed using a differential centrifugation step that separates Golgi membranes and secretory vesicles. This assay will be used to purify and reconstitute proteins required in the budding event. The pathways of vesicle traffic from the yeast Golgi membrane extrapolate to the same event in human cells. As a result of this conservation, yeast cells have been used as a convenient vehicle for the expression and secretion of commercial quantities of therapeutic human proteins. A mechanistic appreciation of the details of vesicle traffic in yeast will also illuminate the pathology of human diseases that impinge on the secretory pathway.

描述（由申请人提供）：蛋白质分泌是一个基本保守的过程，它为细胞创造稳定的细胞内区室（如内质网、高尔基体、溶酶体和质膜）以及与其他细胞和组织交流提供了机制。对酿酒酵母这一过程的分子机制分析揭示了所有真核生物共有的基因和机制。