VTC Events Mediated by the Rab2 Complex

由 Rab2 Complex 主持的 VTC 活动

• 批准号: 7045990
• 负责人: ELLEN J TISDALE
• 金额: $ 24.15万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7045990
• 关键词: Golgi apparatus; SDS polyacrylamide gel electrophoresis; cell line; confocal scanning microscopy; endoplasmic reticulum; enzyme mechanism; glyceraldehyde 3 phosphate dehydrogenase; green fluorescent proteins; guanosinetriphosphatases; isozymes; mass spectrometry; metastasis; microtubules; neoplasm /cancer; pathologic process; protein binding; protein kinase C; protein localization; protein protein interaction; protein structure function; protein transport; proteomics; vesicle /vacuole

DESCRIPTION (provided by applicant): Our long-term goal is to understand the mechanism of protein sorting and recycling from vesicular tubular clusters (VTCs). VTCs function as transport complexes delivering cargo from the ER to the Golgi complex. Moreover, these structures are the first site of segregation of the anterograde and retrograde pathways and thereby perform a critical function. The small GTPase Rab2 plays an essential role in this trafficking step by promoting formation of retrograde-directed vesicles that carry recycling cargo back to the ER. Rab2- mediated vesicle formation requires atypical PKCi/X and GAPDH. In this application we propose to characterize Rab2 mechanistically and to establish the role of Rab2 in events associated with VTCs. The Specific Aims are: 1) to identify the protein components of the VTC subcompartment that binds Rab2, 2) to characterize the interaction between components of the Rab2 complex by mapping their reciprocal binding domains, and 3) elucidate the molecular mechanism of Rab2-dependent-retrograde vesicle budding from VTCs. The importance of investigating Rab2 function in secretion is underscored by the observation that elevated Rab2, PKC, and GAPDH expression correlates with high metastatic potential in various cancers. Since metastasis is a reflection of cytoskeletal reorganization and increased cell motility, it is noteworthy that Rab2 and its effectors influence microtubule dynamics. However, the precise function of Rab2, aPKCi/A, and GAPDH in the early secretory pathway and how these proteins contribute to the disease are unknown. The studies proposed in this application will develop a new and novel paradigm that couples Rab2-Apkc lA-GAPDH dysfunction with pathological consequences.

描述（由申请人提供）：我们的长期目标是了解来自囊泡管状簇（VTC）的蛋白质分类和回收的机制。 VTC的运输配合物的功能从ER到高尔基体配合物。此外，这些结构是顺序和逆行途径的隔离的第一个位置，从而执行关键功能。小型GTPase RAB2通过促进将回收货物回到ER的逆行定向囊泡的形成，在此贩运步骤中起着至关重要的作用。 Rab2-介导的囊泡形成需要非典型的PKCI/X和GAPDH。在此应用程序中，我们建议以机械的方式表征RAB2并确定RAB2在与VTC相关的事件中的作用。具体目的是：1）确定结合rab2，2）VTC子分会的蛋白质成分，通过映射其相互结合结构域来表征Rab2复合物的组成部分之间的相互作用，而3）阐明了Rab2依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性 - 依赖性元素。研究Rab2在分泌中的功能的重要性强调了升高的RAB2，PKC和GAPDH表达与各种癌症中高转移潜力相关的观察结果。由于转移是细胞骨架重组和细胞运动增加的反映，因此值得注意的是，Rab2及其效应子会影响微管动力学。然而，在早期分泌途径中，Rab2，apkci/a和GAPDH的精确功能以及这些蛋白质如何促进该疾病。本应用程序中提出的研究将开发出一种新的新型范式，该范式将Rab2-apkc La-gapdh功能障碍与病理后果相结合。