Rho GTPases and Diaphanous related Formins in HIV-1 replication

HIV-1 复制中的 Rho GTPases 和透明相关福尔明

• 批准号: 45277349
• 负责人: Professor Dr. Oliver T. Fackler, Ph.D.
• 金额: --
• 依托单位: Institut für Experimentelle und Klinische Pharmakologie und Toxikologie
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2007
• 资助国家: 德国
• 起止时间: 2006-12-31 至 2009-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/45277349?language=en
• 关键词: Rho; GTPases; Diaphanous; related; Formins

As obligate cell parasites replication of Human Immunodeficiency Viruses depends on multiple machineries of their target host cells including those regulating cytoskeletal architecture and dynamics. Previous studies implied small Rho GTPases (Rho, Rac and Cdc42) as well as actin and microtubule filament systems in individual steps of the HIV-1 replication cycle. However, the precise functional interactions of HIV-1 with host cell Rho GTPases and their downstream effectors are largely unknown. Our initial goal of this research proposal will be the kinetic and quantitative analysis of Rho GTPase activities over the course of a single round of HIV-1 replication using a synchronized cell system of viral cell-cell transmission. These studies aim at combining biochemical measurements of overall endogenous GTPase activities with imaging approaches to localize activated GTPases in HIV-1 infected cells. Secondly, we will characterize the involvement of cellular machineries that control actin polymerization downstream of small GTPases, such as Diaphanous related Formins or the Arp2/3 complex in HIV-1 replication. These studies will employ HIV permissive cells lines in which expression of individual actin nucleators can be suppressed via Tet-inducible RNAi expression. Components identified to play a role in virus replication will subsequently be analyzed for the localization of their action in the HIV life cycle as well as their underlying molecular mechanism. Together, these studies shall yield a detailed map of the functional interplay of HIV-1 infection and the regulation of host cell small GTPases and their actin polymerizing effectors.

作为专性细胞寄生虫，人类免疫缺陷病毒的复制依赖于其靶宿主细胞的多种机制，包括调节细胞骨架结构和动力学的机制。以前的研究暗示小Rho GTP酶（Rho，Rac和Cdc42）以及肌动蛋白和微管丝系统在HIV-1复制周期的各个步骤。然而，HIV-1与宿主细胞Rho GTP酶及其下游效应物的精确功能相互作用在很大程度上是未知的。我们的初步目标，这项研究的建议将是动力学和定量分析的Rho GTdR活动的过程中的一轮HIV-1的复制使用的病毒细胞-细胞传播的同步细胞系统。这些研究的目的是结合生物化学测量的整体内源性GTP酶活性与成像方法定位激活的GTP酶在HIV-1感染的细胞。其次，我们将描述参与控制肌动蛋白聚合下游的小GTP酶，如透明相关的Formins或Arp2/3复合体在HIV-1复制的细胞机制。这些研究将采用HIV允许细胞系，其中单个肌动蛋白成核剂的表达可以通过Tet-inducible RNAi表达来抑制。确定在病毒复制中发挥作用的组件随后将分析其在HIV生命周期中的作用及其潜在的分子机制。总之，这些研究将产生一个详细的地图的功能相互作用的HIV-1感染和宿主细胞的小GTP酶和它们的肌动蛋白聚合效应的调节。