Role of nucleic acid structure in HIV-1 replication

由人类免疫缺陷病毒1型（HIV-1）引起的获得性免疫缺陷综合征（AIDS）是世纪初化疗面临的最重要挑战之一。在缺乏有效疫苗的情况下，HIV-1逆转录酶和蛋白酶的抑制剂的组合为控制HIV感染提供了强有力的支持。最近，HIV-1整合酶抑制剂也被添加到该药物方案中。然而，开发针对HIV-1的有效疗法的主要挑战是出现了多药耐药病毒株，其包含病毒酶的突变。因此，获得这些关键病毒酶的详细机制信息对于持续开发更好的药物至关重要。 在HIV-1复制期间，RNA基因组通过HIV-1逆转录酶逆转录成整合能力的双链DNA。该病毒DNA随后通过HIV-1整合酶整合到宿主基因组中。我的长期目标是破译在逆转录病毒复制过程中，病毒和细胞酶与其核酸底物之间相互作用的分子细节。 这是基于我的初步和已发表的数据，其中核酸结构的改变 影响它们被HIV-1逆转录酶识别。该提案的核心假设是，核酸的结构在逆转录病毒复制过程中起着重要作用。我的假设的基本原理是基于逆转录病毒酶适应构象不同的核酸底物的方式。利用我的核苷类似物方法和提出的病毒学培训，我将以特定的目的验证我的假设：1）核酸结构/几何形状如何影响HIV-1整合和2）APOBEC 3G如何识别单链病毒DNA。从我的研究中获得的新的和重要的生化数据将有助于我们理解HIV-1复制的机制， 设计出更好、更有效的抗艾滋病药物至关重要。

Acquired immunodeficiency syndrome (AIDS), caused by human immunodeficiency virus type 1 (HlV-1) is one of the most important challenges for chemotherapy of the early 21 st century. In the absence of an effective vaccine, a combination of inhibitors of HlV-1 reverse transcriptase and protease provide strong support for control of HIV infection. Recently, HlV-1 integrase inhibitors have also been added to this drug regimen. However, a major challenge in developing an effective therapy against HIV-1 is the emergence of multidrug-resistant virus strains, containing mutations in viral enzymes. Therefore, it is essential to obtain detailed mechanistic information on these key viral enzymes for the continued development of better drugs. During HlV-1 replication, the RNA genome is reverse transcribed into an integration competent double stranded DNA by HlV-1 reverse transcriptase. This viral DNA is subsequently integrated into the host genome by the HlV-1 integrase. My long term goal is to decipher the molecular details of interactions between essential viral and cellular enzymes with their nucleic acid substrates during retroviral replication. This is based on my preliminary and published data, where alterations in nucleic acid structure have influenced their recognition by HlV-1 reverse transchptase. The central hypothesis of the proposal is that structures of nucleic acids play an important role during retroviral replication. The rationale of my hypothesis is based on the manner in which retroviral enzymes accommodate conformationally-distinct nucleic acid substrates. Exploiting my nucleoside analog methodology and proposed virology training, I will validate my hypothesis with specific aims: 1) How does the nucleic acid structure/geometry influence HlV-1 integration and 2) How APOBEC3G recognizes the single stranded viral DNA. New and important biochemical data obtained from my studies will facilitate our understanding on the mechanism of HlV-1 replication, which is essential to design better and effective drugs against HIV.