Role of p115 in membrane traffic

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。该提案建立在伯明翰亚拉巴马大学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资助的RO 1 GM 62696 -06的延伸，标题为“p115在膜运输中的作用”。通过分析运输因子p115的作用，父授权解决了ER-高尔基体交通的基本机制。P115通过参与蛋白质-蛋白质相互作用与定位在沿着通路的不同隔室中的不同蛋白质在ER-高尔基体运输中起作用。本研究的实验目标是通过分析p115与两种高尔基体蛋白GM 130和giantin的相互作用来探索p115在高尔基体的功能。 现在，我们希望扩大我们的调查范围，探索p115介导的事件前高尔基体阶段的交通。FIRCA的建议是基于我们最近的研究结果，即p115可能在ER出口位点的水平上起作用，产生前高尔基体转运中间体。我们现在计划测试的假设，货物和p115协同招聘ER出口网站和p115协会是必要的，为形成囊泡管状集群（VTC）运输货物和p115到高尔基体。一系列实验方法，包括生物化学，分子，细胞和成像技术将用于解决以下具体目标：1）确定p115如何与ER出口位点相关联; 2）确定p115如何运输到高尔基体;和3）确定p115如何在前高尔基体运输中发挥作用。完成拟议的研究将扩大我们的理解如何p115被招募到膜和移动沿着线性分泌途径，并将提供第一次调查的分子事件，p115介导的运输能力VTC的形成。

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.