Untersuchungen zu den molekularen Mechanismen der Interaktionen zwischen Endoplasmatischem Reticulum und Mikrotubuli unter Verwendung von Vaccinia Virus-Replikations-Komplexen als Modellsystem

以牛痘病毒复制复合体为模型系统研究内质网与微管相互作用的分子机制

• 批准号: 5362171
• 负责人: Dr. Birgit Schramm
• 金额: --
• 依托单位: Europäisches Laboratorium für Molekularbiologie (EMBL)
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2002
• 资助国家: 德国
• 起止时间: 2001-12-31 至 2003-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5362171?language=en
• 关键词: Untersuchungen; zu; den; molekularen; Mechanismen

Although microtubules (MTs) are known to play an important role in structural organization of the endoplasmic reticulum (ER) and ER-dynamics, relatively little is known about the molecular details underlying these interactions. The current proposal aims to use a novel in vitro system, based on mammalian cells, to study the molecular mechanisms of MT/ER interactions. It takes advantage of the observation that the cytoplasmic sites of vaccinia virus (vv) replication become entirely wrapped by the ER and these ER-enclosed sites move in a MT-dependent fashion towards the perinuclear region of the cell. Since the ER-enwrapped replication-sites can be isolated from infected cells, I propose to reconstitute their MT-interactions in vitro. Using a light microscopy assay, I will study first binding to, and then motility along MTs of isolated replication sites. With this assay I expect to gain important new insight into cellular as well as possibly viral factors underlying ER/MT interactions. Finally, the fact that the minus-end directed motility of the replication sites does not depend on the dynactin/dynein complex, suggests that a novel microtubule motor machinery needs to be identified.

虽然微管（MT）是已知的内质网（ER）和ER动力学的结构组织中发挥重要作用，相对知之甚少，这些相互作用的分子细节。目前的建议旨在使用一种新的体外系统，基于哺乳动物细胞，研究MT/ER相互作用的分子机制。它利用了观察到的牛痘病毒（vv）复制的细胞质位点完全被ER包裹，并且这些ER封闭的位点以MT依赖的方式向细胞的核周区域移动。由于ER-β复制位点可以从感染的细胞中分离出来，我建议在体外重建它们的MT相互作用。使用光学显微镜分析，我将首先研究结合，然后沿沿着孤立的复制位点的MT。通过这种检测，我希望获得重要的新的洞察细胞以及可能的病毒因素的基础ER/MT的相互作用。最后，事实上，负端定向运动的复制位点不依赖于动力蛋白/动力蛋白复合物，表明一种新的微管电机机械需要确定。