Role of p115 in membrane traffic

p115 在膜运输中的作用

• 批准号: 6631118
• 负责人: ELIZABETH S SZTUL
• 金额: $ 4.03万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6631118
• 关键词: Golgi apparatus; SDS polyacrylamide gel electrophoresis; South America; binding sites; electron microscopy; endoplasmic reticulum; guanine nucleotide binding protein; immunofluorescence technique; membrane transport proteins; protein binding; protein localization; protein protein interaction; protein structure function; secretory protein; time resolved data; vesicle /vacuole; western blottings

DESCRIPTION (provided by applicant) The goals of this FIRCA application address fundamental cellular processes by exploring molecular mechanisms regulating cargo progression through the secretory pathway. The proposal builds on research interests of collaborating investigators at the University of Alabama at Birmingham and the University of Cordoba, Argentina. The proposal is an extension of a NIH grant RO1 GM62696-06 entitled "Role of p115 in Membrane Traffic". The parent grant addresses basic mechanisms of ER-Golgi traffic by analyzing the role of the transport factor p115. P115 functions in ER-Golgi traffic by participating in protein-proteins interaction with distinct proteins localized in different compartments along the pathway. The experimental goals of the parent grant are to explore p115 function at the Golgi by analyzing tethering interactions of p115 with two Golgi proteins, GM130 and giantin. We now wish to expand the scope of our inquiry by exploring p115-mediated events in pre-Golgi stages of traffic. This FIRCA proposal is based on our recent findings that p115 may act at the level of ER exit sites to generate pre-Golgi transport intermediates. We now plan to test the hypothesis that cargo and p115 are coordinately recruited at ER exit sites and that p115 association is required for the formation of vesicular tubular clusters (VTCs) that transport cargo and p115 to the Golgi. A series of experimental approaches including biochemical, molecular, cellular and imaging technologies will be used to address the following Specific Aims: 1) define how p115 associates with ER exit sites; 2) define how p115 traffics to the Golgi; and 3) define how p115 functions in pre-Golgi transport. Completion of proposed studies will extend our understanding of how p115 is recruited to membranes and moves along the linear secretory pathway, and will provide first ever inquiry into the molecular events by which p115 mediates the formation of transport competent VTCs.

描述（由申请人提供） 该 FIRCA 应用的目标是通过探索通过分泌途径调节货物进展的分子机制来解决基本的细胞过程。该提案建立在阿拉巴马大学伯明翰分校和阿根廷科尔多瓦大学合作研究人员的研究兴趣之上。该提案是 NIH 拨款 RO1 GM62696-06 题为“p115 在膜交通中的作用”的延伸。父资助通过分析运输因子 p115 的作用来解决 ER-高尔基体运输的基本机制。 P115 通过参与与位于沿通路不同区室的不同蛋白质的蛋白质-蛋白质相互作用，在 ER-高尔基体运输中发挥作用。父母资助的实验目标是通过分析 p115 与两种高尔基体蛋白 GM130 和 Giantin 的束缚相互作用来探索 p115 在高尔基体的功能。 我们现在希望通过探索前高尔基体阶段的 p115 介导的事件来扩大我们的研究范围。该 FIRCA 提案基于我们最近的发现，即 p115 可能在 ER 出口位点水平发挥作用，生成前高尔基体转运中间体。我们现在计划检验以下假设：货物和 p115 在 ER 出口位点协调招募，并且 p115 关联是形成将货物和 p115 运输到高尔基体的囊泡管状簇 (VTC) 所必需的。将使用包括生化、分子、细胞和成像技术在内的一系列实验方法来实现以下具体目标：1）定义p115如何与ER出口位点结合； 2) 定义p115如何到达高尔基体； 3) 定义 p115 在前高尔基体运输中的功能。拟议研究的完成将扩展我们对 p115 如何被招募到膜上并沿着线性分泌途径移动的理解，并将首次探究 p115 介导具有运输能力的 VTC 形成的分子事件。