Drs2p Function in Clathrin-coated Vesicle Budding

Drs2p 在网格蛋白包被的囊泡出芽中的功能

• 批准号: 6655530
• 负责人: TODD R GRAHAM
• 金额: $ 24.16万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-15 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6655530
• 关键词: ADP ribosylation; G protein; Saccharomyces cerevisiae; cell component structure /function; cell free system; clathrin; electron microscopy; fungal genetics; fungal proteins; hydrogen potassium exchanging ATPase; intracellular transport; membrane lipids; membrane reconstitution /synthesis; membrane transport proteins; molecular site; phospholipids; posttranslational modifications; protein localization; protein structure function; site directed mutagenesis; technology /technique development; vesicle /vacuole; ADP ribosylation; G protein; Saccharomyces cerevisiae; cell component structure /function; cell free system; clathrin; electron microscopy; fungal genetics; fungal proteins; hydrogen potassium exchanging ATPase; intracellular transport; membrane lipids; membrane reconstitution /synthesis; membrane transport proteins; molecular site; phospholipids; posttranslational modifications; protein localization; protein structure function; site directed mutagenesis; technology /technique development; vesicle /vacuole

DESCRIPTION (APPLICANT'S ABSTRACT): A striking feature of eukaryotic cells, from yeast to human, is their compartmental organization into membrane-bound organelles, each with a unique composition and function. The long-term goal of this research project is to understand how the Goigi complex, a multi-compartment organelle, performs its essential role in the transport, modification and sorting of proteins in the secretory pathway. Towards this goal, the function of ADP-ribosylation factor (ARF) in vesicle-mediated transport from the Golgi complex is being examined. ARF appears to regulate the budding of both COPI- and clathrin-coated vesicles from the Golgi complex. The proposed studies stem from recent discoveries implicating a role for Drs2p, an integral membrane P-type ATPase and potential lipid translocase (or flippase), specifically in the ARF-dependent budding of clathrin coated vesicles. The yeast Saccharomyces cerevisiae will be used for this work because of the fundamental similarity of the secretory pathway in all eukaryotic cells coupled with the powerful genetic and molecular approaches available with this organism make it an ideal model system. The specific goals of this research project are to determine if Drs2p is required for recruiting ARF and clathrin to Golgi membranes, thus indicating a direct role for Drs2p in clathrin coated vesicle budding. The proposed lipid flippase activity of Drs2p will be tested as will the potential overlap in function between Drs2p and four Drs2p-related proteins encoded within the yeast genome. There are profound implications to human health that can be derived from this study. Both Drs2p and clathrin are required for the normal function of the Golgi complex in the transport and processing of many proteins. In addition, lipid flippases related to Drs2p are thought to restrict phosphatidylserine (PS) to the inner leaflet of 'the plasma membrane. Loss of this asymmetric distribution of PS in blood cells stimulates clot formation and in apoptotic cells allows for recognition and phagocytosis by macrophages.

描述（申请人的摘要）：真核细胞的引人注目的特征， 从酵母到人，是他们的隔室组织到膜的结合 细胞器，每个细胞器具有独特的组成和功能。长期目标 该研究项目是为了了解Goigi Complex如何 多室内细胞器在运输中发挥重要作用， 分泌途径中蛋白质的修改和分类。走向这个 目标，囊泡介导的ADP-核糖基化因子（ARF）的功能 正在研究从高尔基体综合体的运输。 ARF似乎调节了 来自高尔基体复合物的辅料和网格蛋白涂层囊泡的萌芽。这 拟议的研究源于最近的发现，暗示了DRS2P的作用 整体膜P型ATPase和潜在的脂质易位酶（或Flippase）， 特别是在网状蛋白涂层囊泡的ARF依赖性萌芽中。这 酿酒酵母将用于这项工作，因为 所有真核细胞中分泌途径的基本相似性耦合 使用强大的遗传和分子方法可用 使其成为理想的模型系统。该研究项目的具体目标是 确定是否需要DRS2P招募ARF和Clathrin到Golgi 膜，因此表明drs2p在网格蛋白涂层囊泡中的直接作用 萌芽。 drs2p的拟议的脂质氟夫酶活性将被测试 DRS2P和四个DRS2P相关蛋白之间功能的潜在重叠 编码在酵母基因组中。对人有深远的影响 可以从这项研究中得出的健康。 Drs2p和clathrin都是 高尔基综合体在运输中的正常功能所必需的 处理许多蛋白质。此外，与DRS2P相关的脂质翻盘是 被认为将磷脂酰丝氨酸（PS）限制为血浆的内部小叶 膜。血细胞中PS的这种不对称分布的损失刺激 凝块形成和凋亡细胞中可以识别和吞噬作用 通过巨噬细胞。