Mechanism and Regulation of HIV-1 Uncoating

描述（由申请方提供）：HIV-1感染是通过病毒和靶细胞膜融合引发的，导致核糖核蛋白核心递送到宿主细胞质中。对于所有逆转录病毒，融合后立即发生的事件知之甚少。通常认为病毒核心经历了涉及病毒衣壳溶解的脱壳过程。我们实验室的研究表明，HIV-1衣壳稳定性的相对细微的扰动导致感染性差的病毒粒子在靶细胞中的病毒DNA合成受损，这表明脱壳是一个微调的过程，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敏感性，以及限制因子TRIM 51通过靶向CA阻断HIV-1感染的能力，表明去包被可能是抗病毒治疗的有吸引力的靶点。除了CA，其他病毒和细胞分子可能参与调节HIV-1在体内的脱壳。在本申请中，我们建议使用生物化学和基于细胞的检测，以更好地了解HIV-1的脱壳机制，并确定相关的病毒-宿主细胞相互作用，适合治疗靶向。这项工作将根据五个具体目标进行规划：1。分析靶细胞中感染性差的HIV-1 CA突变体的脱壳。2.鉴定HIV-1非包被突变体的新型假回复突变体。3.鉴定HIV-1脱壳过程中的结构中间体。4.鉴定调节HIV-1脱壳的宿主细胞因子。5.研究TRIM 51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。 公共卫生相关性：HIV-1感染严重依赖于对病毒核心的脱壳过程知之甚少。我们将进行生物化学和分子遗传学研究，以确定病毒和细胞的决定因素的脱壳，并确定在这个过程中的阶段。最终，这些研究将有助于确定在开发新的抗逆转录病毒疗法时，脱膜是否可以成为一个有用的靶点。

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.