HIV-1 Integrase Structural Biology

描述(由申请人提供)：整合由病毒整合酶蛋白催化，是所有逆转录病毒生命周期中的重要步骤。这突出了人类免疫缺陷病毒1型(HIV-1)整合酶蛋白是治疗干预的重要目标，第一种整合酶抑制剂Raltegravir于2007年被批准用于艾滋病患者。由于患者对拉替格列韦产生耐药性，因此需要不断开发下一代抑制剂，以对抗耐药菌株，这项工作正在制药业进行。对靶蛋白的详细结构以及药物结合复合体的结构的了解极大地有助于此类药物设计工作。与游离整合酶蛋白相比，雷替格列韦等药物优先结合并抑制逆转录后形成的整合酶-DNA复合体或整合酶小体，突显了对HIV-1整合酶小体结构信息的迫切需要。直到最近，还没有任何逆转录病毒内切体的实验衍生结构，但在正在进行的赠款平台期间进行的工作最终解决了泡沫病毒(PFV)原型内切酶的X射线晶体结构。为了开始了解类似的HIV-1结构，我们建立了一个以PFV结构为支架的分子模型，本文提出的工作将评估模型中揭示的许多整合酶-DNA接触在体外整合和病毒感染过程中的作用。此外，我们将采用我们在对PFV进行广泛研究时所学到的知识，并将这些知识应用于解决HIV-1连接体的三维结构。HIV-1结构将是基础研究和临床科学家的宝贵财富，因为它将定义HIV-1 DNA整合的结构基础，并为开发下一代整合酶抑制剂提供关键平台。

DESCRIPTION (provided by applicant): Integration, catalyzed by the viral integrase protein, is an essential step in the life cycle of all retroviruses. This has highlighted the human immunodeficiency virus type 1 (HIV- 1) integrase protein as an important target for therapeutic intervention, and the first integrase inhibitor, raltegravir, was approved for use with AIDS patients in 2007. Because raltegravir-resistance arises in patients, there is an ongoing, important need to develop next generation inhibitors that will work to counteract drug-resistant strains, and this work is underway in the pharmaceutical industry. Such drug design efforts are helped significantly by knowledge of detailed structures of the targeted protein, as well as structures of drug-bound complexes. Drugs like raltegravir preferentially bind to and inhibit the integrase-DNA complex or intasome that forms after reverse transcription as compared to the free integrase protein, highlighting the crucial need for structural information on the HIV-1 intasome. Until recently, there was no experimentally derived structure for any retroviral intasome, but work conducted during the ongoing grant platform culminated in solving the x-ray crystal structure of the prototype foamy virus (PFV) intasome. To begin to understand the analogous HIV-1 structure, we have built a molecular model using the PFV structure as a scaffold, and work proposed herein will evaluate numerous integrase-DNA contacts unveiled in the model for their roles in integration in vitro and during virus infection. We moreover will adopt what we have learned during our extensive studies with PFV and apply this knowledge to solve the three-dimensional structure of the HIV-1 intasome. The HIV-1 structure will be an invaluable asset for basic research as well as clinical scientists, as it will define the structural basis of HIV-1 DNA integration and provide a crucial platform for development of next generation integrase inhibitors.