MOLECULAR INTERACTIONS OF NEF AND AIDS PATHOGENESIS

描述：(改编自申请者摘要)这项工作的总体目标 应用是理解Nef和Nef之间的分子相互作用 Nef在艾滋病发病机制中发挥作用所必需的细胞蛋白。 分子遗传学和生物化学方法将被用来解剖几个 HIV-1和SIV Nef的功能，包括Nef诱导 CD4和MHC I类分子的内吞作用，以改变细胞信号 转导途径，并加强病毒复制。这些实验 将揭示Nef和Nef组分之间的多分子相互作用 作为其基础的细胞信号转导和内吞机制 功能。Nef的这些分子相互作用对艾滋病的重要性 发病机制将使用SIV感染的恒河猴模型来解决 在对直接从HIV-1感染中分离的天然Nef蛋白的研究中 人民。具体地说，前两组实验将表征 Nef与CD4和MHC类分子胞质结构域的相互作用 I分子使用遗传和生化技术并将识别 分离个体功能和/或分子的Nef突变 Nef与胞内和信号成分的相互作用 换能器。第三组实验将使用酵母 双杂交系统用于鉴定介导功能的细胞蛋白 Nef与内吞和信号转导机制的相互作用。 最后，他们将研究突变对所选基因的影响 Nef在恒河猴艾滋病发病中的分子相互作用。这些 研究将为理解这些功能提供一个分子框架 Nef，这些功能及其背后的分子相互作用 对致病过程的重要性。他们的研究结果将会有 对合理设计有效战略的影响 艾滋病所需的Nef功能的药理学干扰 发病机制。

DESCRIPTION: (Adapted from Applicant's Abstract) The overall goal of this application is to understand the molecular interactions between Nef and the cellular proteins essential for the role of Nef in AIDS pathogenesis. Molecular-genetic and biochemical approaches will be used to dissect several functions of HIV-1 and SIV Nef, including the ability of Nef to induce endocytosis of CD4 and MHC class I molecules, to alter cellular signal transduction pathways, and to enhance viral replication. These experiments will uncover multiple molecular interactions between Nef and components of the cellular signal transduction and endocytic machinery that underlie these functions. The importance of these molecular interactions of Nef for AIDS pathogenesis will be addressed using the SIV-infected rhesus monkey model and in studies of natural Nef proteins isolated directly from HIV-1 infected people. Specifically, the first two sets of experiments will characterize the interaction between Nef and the cytoplasmic domains of CD4 and MHC class I molecules using genetic and biochemical techniques and will identify mutations in Nef that dissociate individual functions and/or molecular interactions between Nef and the components of endocytic and signal transduction machineries. The third set of experiments will use the yeast two-hybrid system to identify cellular proteins that mediate functional interactions of Nef with the endocytic and signal transduction machineries. Finally, they will study the effect of mutations disrupting selected molecular interactions of Nef on AIDS pathogenesis in rhesus monkeys. These studies will provide a molecular framework for understanding the functions of Nef, the molecular interactions that underlie these functions and their importance for the pathogenic process. Results from their studies will have implications for a rational design of effective strategies for pharmacological disruption of Nef functions that are required for AIDS pathogenesis.