Mechanism and Regulation of HIV-1 Uncoating

描述(申请人提供)：HIV-1感染是通过病毒和目标细胞膜的融合启动的，导致核糖核蛋白核心进入宿主细胞细胞质。对于所有逆转录病毒来说，融合后紧接着发生的事件知之甚少。通常认为，病毒核心经历了一个涉及病毒衣壳溶解的剥离过程。我们实验室的研究表明，HIV-1衣壳稳定性的相对微小扰动会导致感染性较差的病毒粒子，从而损害靶细胞中病毒DNA的合成，这表明去涂层是一个微调的过程，对于高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来阻断HIV-1感染的能力，表明去涂层可能是抗病毒治疗的一个有吸引力的靶点。除了CA，其他病毒和细胞分子也可能参与体内HIV-1脱壳的调节。在这一应用中，我们建议使用生化和基于细胞的分析来更好地了解HIV-1去涂层的机制，并确定适合于治疗靶向的相关病毒-宿主细胞相互作用。这项工作将按照五个具体目标进行计划：1.分析低感染性HIV-1 CA突变体在靶细胞中的脱壳情况。2.鉴定新的HIV-1脱壳突变体的假复原体。3.确定HIV-1去涂层中的结构中间体。4.确定调节HIV-1脱壳的宿主细胞因子。5.体外检测TRIM51对HIV-1去包被的影响。总而言之，这些研究将阐明艾滋病毒-1生命周期中一个模糊的阶段，并揭示艾滋病毒/艾滋病治疗的新可能性。 公共卫生相关性：HIV-1感染严重依赖于鲜为人知的揭开病毒核心的过程。我们将进行生化和分子遗传学研究，以确定脱壳的病毒和细胞决定因素，并确定过程中的各个阶段。最终，这些研究将有助于确定去涂层是否可以成为开发新的抗逆转录病毒疗法的有用靶点。

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.