Mechanism and Regulation of HIV-1 Uncoating

描述（由申请人提供）：HIV-1感染是由病毒与靶细胞膜融合引起的，导致核糖核蛋白核心进入宿主细胞质。对于所有逆转录病毒，融合后立即发生的事件了解甚少。一般认为，病毒核心经历了一个涉及病毒衣壳溶解的剥膜过程。我们实验室的研究表明，HIV-1衣壳稳定性相对微小的扰动导致感染性较差的病毒粒子在靶细胞中的病毒DNA合成受损，这表明脱衣是一个精细调整的过程，对有效的逆转录至关重要。HIV-1感染对衣壳稳定性改变的高度敏感性，以及限制因子TRIM51通过靶向CA阻断HIV-1感染的能力，表明脱膜可能是抗病毒治疗的一个有吸引力的靶点。除CA外，其他病毒分子和细胞分子也可能参与体内HIV-1脱壳的调控。在这个应用中，我们建议使用生化和基于细胞的分析来更好地了解HIV-1脱膜的机制，并确定适合治疗靶向的相关病毒-宿主细胞相互作用。这项工作将根据五个具体目标进行规划：1。目的分析低感染性HIV-1 CA突变体在靶细胞中的脱壳。2. 鉴定HIV-1脱包衣突变体的新型假逆转物。3. 鉴定HIV-1脱包衣的结构中间体。4. 鉴定调控HIV-1脱膜的宿主细胞因子。5. 探讨TRIM51对HIV-1体外脱膜的影响。总的来说，这些研究将阐明HIV-1生命周期的一个模糊阶段，并揭示HIV/AIDS治疗的新可能性。

DESCRIPTION (provided by applicant): HIV-1 infection is initiated by fusion of virus and target cell membranes, resulting in delivery of the ribonucleoprotein core into the host cell cytoplasm. The ensuing events that immediately follow fusion are poorly understood for all retroviruses. It is generally assumed that the viral core undergoes an uncoating process involving dissolution of the viral capsid. Studies in our laboratory have shown that relatively subtle perturbations in HIV-1 capsid stability result in poorly infectious virions that are impaired for viral DNA synthesis in target cells, suggesting that uncoating is a finely tuned process and is crucial for efficient reverse transcription. The high sensitivity of HIV-1 infection to alterations in capsid stability, and the ability of the restriction factor TRIM51 to block HIV-1 infection by targeting CA, suggest that uncoating may be an attractive target for antiviral therapy. Besides CA, other viral and cellular molecules are likely to participate in the regulation of HIV-1 uncoating in vivo. In this application, we propose to use biochemical and cell-based assays to better understand the mechanism of HIV-1 uncoating and to identify relevant virus-host cell interactions suitable for therapeutic targeting. The work will be planned according to five Specific Aims: 1. To analyze the uncoating of poorly infectious HIV-1 CA mutants in target cells. 2. To identify novel pseudorevertants of HIV-1 uncoating mutants. 3. To identify structural intermediates in HIV-1 uncoating. 4. To identify host cell factors which regulate HIV-1 uncoating. 5. To determine the effect of TRIM51 on HIV-1 uncoating in vitro. Collectively, these studies will elucidate an obscure stage of the HIV-1 life cycle and reveal new possibilities for HIV/AIDS therapy. PUBLIC HEALTH RELEVANCE: HIV-1 infection is critically dependent on the poorly understood process of uncoating of the viral core. We will perform biochemical and molecular genetic studies to identify viral and cellular determinants of uncoating and to define stages in the process. Ultimately, these studies will help determine whether uncoating can be a useful target in the development of new antiretroviral therapies.