Role of p115 in membrane traffic

描述(由申请人提供) FIRCA应用的目标是通过探索通过分泌途径调节货物进程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基础上。该提案是美国国立卫生研究院题为“p115在膜交通中的作用”的RO1 GM62696-06拨款的延伸。父授权通过分析传输因子p115的作用来解决ER-Golgi业务的基本机制。P115在内质网-高尔基体运输中的作用是通过参与不同蛋白质之间的相互作用，这些不同的蛋白质位于不同的通道中。亲本基金的实验目标是通过分析p115与两种高尔基蛋白GM130和giantin的拴系相互作用来探索p115在高尔基体中的功能。 我们现在希望通过探索在交通的前高尔基阶段中由p115介导的事件来扩大我们的调查范围。FIRCA的这一建议是基于我们最近的发现，即p115可能在内质网出口部位水平起作用，产生高尔基体前运输中间产物。我们现在计划检验这样的假设，即货物和p115是在内质网出口部位协同招募的，并且p115的结合是形成将货物和p115运输到高尔基体的囊泡管状团(VTC)所必需的。包括生化、分子、细胞和成像技术在内的一系列实验方法将被用来解决以下具体目标：1)确定p115如何与内质网出口部位相关联；2)确定p115如何向高尔基体传递；3)定义p115在高尔基体前运输中如何发挥作用。这些研究的完成将扩大我们对p115如何被招募到膜上并沿着线性分泌途径移动的理解，并将首次探讨p115介导具有运输能力的VTC形成的分子事件。

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.