Mechanisms and Consequences of Reverse Transcription in HIV-1 Cores

摘要 HIV-1感染会导致T辅助细胞的破坏，导致免疫缺陷和 被称为艾滋病的疾病。目前大多数可用的抗逆转录病毒药物都是针对病毒逆转的 转录酶、整合酶和蛋白水解酶。这些药物是有效的，但不能治愈， 治疗必须严格和永久地坚持下去。此外，HIV-1病毒不断进化 并获得突变，使其对治疗产生抵抗力。因此，需要进行持续的研究。 开发针对新的病毒和细胞靶点的药物。由于它在感染中具有多重功能， HIV-1衣壳代表了一个有吸引力的治疗靶点，以及针对这一点的有效抑制剂 结构在临床试验中显示出良好的效果。衣壳，这是一个组装的格子 由单一病毒蛋白组成，在病毒基因组和相关蛋白周围形成一层外壳 对于高效的逆转录是至关重要的，逆转录是HIV-1感染的关键早期步骤。 对衣壳稳定性的扰动会导致流产感染，但其机制 衣壳确保了有效的逆转录是未知的。该项目将通过以下方式填补这一空白 确定衣壳扰动对HIV-1体外逆转录的影响，反之亦然。 使用从具有传染性的HIV-1颗粒中提纯的天然病毒核心，我们将使用尖端技术 生物物理技术与计算和分子病毒学方法相结合 准确定义病毒衣壳在HIV-1反转录中的作用。该项目将 完成以下目标：(1)确定反转录的生化要求 在HIV-1核心中；(2)确定反转录过程中核心的结构转变；(3) 确定衣壳毛孔在反转录过程中核苷酸摄取中的作用；以及(4) 精确定义在特定阶段核心内的蛋白质-核酸相互作用 逆转录。总的来说，这个项目将对衣壳蛋白的功能有更好的理解 在HIV-1感染中，从而告知一种新兴的抗病毒药物的作用机制 毒品。 相关性 这个项目的结果将是更好地了解病毒衣壳的功能， 对HIV-1感染至关重要。这些信息将为衣壳病毒靶向的发展提供信息 抗病毒药物和改善艾滋病毒感染者的治疗，从而改善公共卫生 以及减少HIV-1的传播。

Summary HIV-1 infection results in destruction of T helper cells, leading to immunodeficiency and the disease known as AIDS. Most currently available antiretroviral drugs target the viral reverse transcriptase, integrase, and protease enzymes. These medicines are effective but not curative, and therapy must be adhered to rigorously and permanently. Moreover, HIV-1 constantly evolves and acquires mutations rendering it resistant to therapies. Therefore, ongoing research is needed to develop drugs against new viral and cellular targets. Owing to its multiple functions in infection, the HIV-1 capsid represents an attractive therapeutic target, and potent inhibitors targeting this structure have shown promising results in clinical trials. The capsid, which is an assembled lattice composed of a single viral protein, forms a shell around the viral genome and associated proteins and is essential for efficient reverse transcription, a key early step in HIV-1 infection. Perturbations to the stability of the capsid result in abortive infection, yet the mechanism by which the capsid ensures efficient reverse transcription is unknown. This project will fill this gap by defining the effects of capsid perturbations on HIV-1 reverse transcription in vitro, and vice-versa. Using native viral cores purified from infectious HIV-1 particles, we will employ cutting-edge biophysical techniques together with computational and molecular virology approaches to precisely define the role of the viral capsid in HIV-1 reverse transcription. The project will accomplish the following goals: (1) define the biochemical requirements for reverse transcription in HIV-1 cores; (2) define the structural transitions in the core during reverse transcription; (3) determine the role of capsid pores in nucleotide uptake during reverse transcription; and (4) precisely define the protein-nucleic acid interactions within the core during specific stages of reverse transcription. Collectively, this project will yield a greater understanding of capsid function in HIV-1 infection, thereby informing the mechanism of action of an emerging class of antiviral drugs. Relevance The results of this project will be a greater understanding of the functions of the viral capsid that are critical for HIV-1 infection. This information will inform the development of capsid-targeting antiviral drugs and improve the treatment of HIV-infected persons, thereby improving public health and reducing the spread of HIV-1.