HIV-1 reverse transcriptase structure: function, inhibition, and resistance

描述（由申请人提供）：HIV-1逆转录酶（RT）是艾滋病抗病毒治疗的中心靶点，对其结构和功能的详细了解具有重要的临床和生物学意义。核苷类和非核苷类逆转录酶抑制剂都被用作治疗艾滋病的有效药物，但耐药病毒变异体的出现可能会限制其成功。通过基于结构的方法开发了新型非核苷类RT抑制剂，包括最近批准的药物etravirine/TMC 125/Intelence，我们建议进行晶体学研究，以实现结构指导的其他类别的RT抑制剂的改进。拟议的工作将涉及在存在和不存在核酸底物和抑制剂的情况下确定野生型和耐药HIV-1 RT的结构。抑制剂的结构研究将探索已知药物（NRTI和NNRTI）的结合位点以及其他潜在的药物靶向位点，包括RNase H活性位点。使用策略性修饰的蛋白质和核酸将提高结构研究的效率和质量。所确定的结构将增强我们对聚合酶和RNase H催化、抑制和耐药性机制的理解。

DESCRIPTION (provided by applicant): HIV-1 reverse transcriptase (RT) is a central target for antiviral treatment of AIDS and detailed knowledge of its structure and function has important clinical and biological consequences. Both nucleoside and non-nucleoside RT inhibitors are used as effective drugs for treating AIDS, but success can be limited by the emergence of drug-resistant viral variants. Novel non-nucleoside RT inhibitors have been developed through structure-based methods, including the recently approved drug etravirine/TMC125/Intelence, and we propose to carry out crystallographic studies that could enable structure-guided improvement of additional classes of RT inhibitors. The proposed work will involve structure determination of wild-type and drug-resistant HIV-1 RT in the presence and absence of nucleic acid substrates and inhibitors. Structural studies of inhibitor will explore binding sites for known drugs (NRTIs and NNRTIs) as well as other potential drug targeting sites including the RNase H active site. Use of strategically modified protein and nucleic acids will enhance the efficiency and quality of the structural studies. The structures determined will enhance our understanding of mechanisms of polymerase and RNase H catalysis, inhibition, and of drug resistance.