NEF AND HIV AND SIV MOLECULAR PATHOGENESIS

NEF 与 HIV 和 SIV 的分子发病机制

• 批准号: 2442673
• 负责人: EARL T SAWAI
• 金额: $ 8.72万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2442673
• 关键词: AIDS; CD4 molecule; Macaca mulatta; T lymphocyte; affinity chromatography; human immunodeficiency virus; leukocyte activation /transformation; molecular pathology; protein kinase; protein structure function; simian immunodeficiency virus; site directed mutagenesis; tissue /cell culture; virus infection mechanism; virus protein

The objective of this First proposal is to elucidate the role of Nef in HIV pathogenesis. A combined molecular genetic, biochemical, and virologic approach will be used to de fine the cellular interactions and biologic function of Nef and to ascertain whether these properties are critical for viral pathogenesis. Hypothesis; The interaction between the Nefs of HIV and SIV and cellular factors, including a newly recognized protein kinase, are critical for the maintenance of high viral loads and disease progression. Specific Aim 1: To elucidate the mechanism of Nef action, it is important to identify the cellular proteins that interact with Nef. Affinity chromatography will be used to identify and characterize the cellular proteins that associate with Nef; emphasis will be directed towards determining the identity and properties of the Nef-associated protein kinase . Specific Aim 2: To define the functional domain(s) of Nef and identify critical determinants for the interactions between Nef and cellular proteins; site-specific Nef mutants will be constructed and tested for kinase association, enhancement of viral infectivity, T cell activation, and CD4 down-regulation. Specific Aim 3: To define the role(s) of subcellular localization and membrane targeting in determining the interactions between Nef and its intracellular targets, and to determine whether the partitioning of Nef into specific subcellular compartments affects its phenotypic properties. Specific Aim 4: To determine whether the interaction of Nef with the cellular protein kinase activity is essential for viral persistence and progression to AIDS in vivo, by using SIV infection of rhesus macaques. Significance: The global spread of HIV infection and AIDS continues while current therapies are very limited and effective vaccines are nonexistent. Accordingly, novel therapeutic approaches must be devised to target viral gene functions that regulate virus replication nd produce disease. Thus, knowledge of Nef function will not only aid in elucidating the mechanism of viral pathogenesis, but also may be used to design novel therapies that inhibit virus replication and block progression to AIDS.

第一个建议的目的是阐明NEF在 HIV发病机理。 分子遗传，生化和 病毒学方法将用于罚款细胞相互作用和 NEF的生物学功能并确定这些特性是否是 对于病毒发病机理至关重要。 假设; 之间的相互作用 HIV和SIV的NEF和细胞因素，包括新认识的 蛋白激酶，对于维持高病毒载荷和 疾病进展。 特定目的1：为了阐明NEF作用的机制，这很重要 确定与NEF相互作用的细胞蛋白。 亲和力 色谱法将用于识别和表征细胞 与NEF关联的蛋白质；重点将指向 确定与NEF相关蛋白的身份和特性 激酶。 特定目的2：定义NEF的功能域并识别 NEF与细胞之间相互作用的关键决定因素 蛋白质；将构建并测试特定地点的NEF突变体 激酶关联，病毒感染性的增强，T细胞活化， 和CD4下调。 特定目的3：定义亚细胞定位的作用和 膜靶向确定NEF及其ITS之间的相互作用 细胞内靶标，并确定NEF的分区是否 进入特定的亚细胞隔室会影响其表型特性。 特定目的4：确定NEF的相互作用是否与 细胞蛋白激酶活性对于病毒持久性和 通过使用恒河猕猴的SIV感染，向体内进展。 意义：艾滋病毒感染和艾滋病的全球蔓延仍在继续 当前的疗法非常有限，有效疫苗不存在。 因此，必须设计新的治疗方法以靶向病毒 调节病毒复制和产生疾病的基因功能。 因此， NEF功能的知识不仅有助于阐明机制 病毒发病机理，但也可以用于设计新的疗法 抑制病毒复制并阻止对艾滋病的进展。