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: 6525985
• 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/6525985
• 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

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（申请人摘要）：真核细胞的一个显着特征， 从酵母到人类，它们的区室组织是膜结合的 细胞器，每个都有独特的组成和功能。长期目标是 这个研究项目是为了了解 Goigi 复合体（Goigi 复合体）是如何 多室细胞器，在运输中发挥重要作用， 分泌途径中蛋白质的修饰和分类。朝这个方向 目标，ADP-核糖基化因子（ARF）在囊泡介导中的功能 正在检查高尔基复合体的运输。 ARF 似乎监管 COPI 和网格蛋白包被的囊泡从高尔基复合体中出芽。这 拟议的研究源于最近的发现，表明 Drs2p 的作用，Drs2p 是一种 整合膜 P 型 ATP 酶和潜在的脂质转位酶（或翻转酶）， 特别是在网格蛋白包被的囊泡的 ARF 依赖性出芽中。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 利用该生物体可用的强大遗传和分子方法 使其成为理想的模型系统。该研究项目的具体目标是 确定将 ARF 和网格蛋白招募到高尔基体是否需要 Drs2p 膜，从而表明 Drs2p 在网格蛋白包被的囊泡中具有直接作用 正在萌芽。 Drs2p 的拟议脂质翻转酶活性将进行测试 Drs2p 和四种 Drs2p 相关蛋白之间的潜在功能重叠 编码在酵母基因组内。对人类有着深远的影响 可以从这项研究中得出的健康状况。 Drs2p 和网格蛋白都是 高尔基复合体在运输和运输中的正常功能所需 许多蛋白质的加工。此外，与 Drs2p 相关的脂质翻转酶是 被认为将磷脂酰丝氨酸 (PS) 限制在“血浆”的内层 膜。血细胞中 PS 不对称分布的丧失会刺激 凝块形成和凋亡细胞中允许识别和吞噬作用 由巨噬细胞。