MOLECULAR STUDIES OF EUKARYOTIC CELL SURFACE GROWTH

真核细胞表面生长的分子研究

• 批准号: 6018507
• 负责人: RANDY W. SCHEKMAN
• 金额: $ 16.98万
• 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 2002-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6018507
• 关键词: 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 transport; secretory protein; vesicle /vacuole; western blottings

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 external cells and tissues. A genetic, molecular cloning, and biochemical analysis of this process in Saccharomyces cerevisiae has uncovered genes and mechanisms that are shared among all eukaryotes. This proposal develops a new line of investigation concerning the mechanism of transport vesicle formation from the trans Golgi cisterna and of the pathway of vesicle traffic to the cytokinesis furrow which forms early in the yeast cell cycle. Evidence has emerged to suggest multiple paths of transport vesicle formation from the trans Golgi, at least one of which involves vesicular traffic through the endosome en route to the cell surface. Genetic and cell fractionation analysis is proposed to relate secretion to the known pathway of sorting to the endosome, to discover new genes involved in budding from the Golgi, and to use biochemical analysis to clarify the role of clathrin in one of the budding pathways. Certain membrane enzymes are transported through the endosome to the nascent division septum where they participate in forming a cell wall structure that marks the site of yeast bud emergence (the chitin ring) early in the cell cycle. The pathway and regulation of transport of these enzymes from the endosome to the nascent septeum will be examined to understand the mechanism and cell cycle control of this essential process. The study of these processes in yeast cells has illuminated fundamental aspects of cell and organelle growth that are shared by human cells. Furthermore, the application of this knowledge has allowed biotechnology and pharmaceutical companies to harness yeast cells to express and secrete commercial quantities of important hormones, blood proteins, growth factors, and viral antigens.

蛋白质分泌是一个基本上保守的过程， 细胞产生稳定的细胞内区室的机制， 内质网，高尔基体，溶酶体和质膜，并与 外部细胞和组织。 一个基因，分子克隆， 对酿酒酵母中这一过程的生化分析， 发现了所有真核生物共有的基因和机制。 这一建议开辟了一条新的调查路线， 跨高尔基池运输囊泡形成机制 以及囊泡运输到胞质分裂沟的途径， 在酵母细胞周期的早期形成。 有证据表明，运输囊泡的多种途径 从transGolgi形成，其中至少有一个涉及囊泡 通过内体到达细胞表面的运输。 遗传和 提出细胞分级分离分析将分泌与已知的 内体的分选途径，以发现参与 从高尔基体出芽，并使用生化分析，以澄清 网格蛋白在其中一个出芽途径中的作用。 某些膜酶通过核内体转运到膜上， 新生分裂隔膜，在那里它们参与形成细胞壁 标志着酵母芽出现的结构（几丁质环） 在细胞周期的早期。 转运的途径和调控 这些酶从核内体到新生隔将被检测 了解这一重要的机制和细胞周期控制， 过程 在酵母细胞中对这些过程的研究阐明了基本的 细胞和细胞器生长的各个方面，这是人类细胞共有的。 此外，这些知识的应用使生物技术 和制药公司利用酵母细胞来表达和 分泌大量的重要激素，血液蛋白， 生长因子和病毒抗原。