Integrase Structural Virology

整合结构病毒学

• 批准号: 9440913
• 负责人: Alan N. Engelman
• 金额: $ 53.32万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9440913
• 关键词: 3-Dimensional; Acquired Immunodeficiency Syndrome; Active Sites; Acute; Anti-HIV Agents; Architecture; Betaretrovirus; Biochemical Genetics; Biological; C-terminal; Centers for Disease Control and Prevention (U.S.); Chemical Warfare; Cleaved cell; Clinical; Complex; Cryoelectron Microscopy; DNA; DNA Binding; DNA Integration; Data; Defect; Development; Dinucleoside Phosphates; Drug Design; Drug Targeting; Enzymes; Funding; Genetic Techniques; Grant; HIV; HIV-1; Highly Active Antiretroviral Therapy; Hydroxyl Radical; In Vitro; Infection; Integrase; Integrase Inhibitors; Life Cycle Stages; Maps; Modeling; Morphogenesis; Mouse Mammary Tumor Virus; Nucleoproteins; Patients; Pharmaceutical Preparations; Phase; Positioning Attribute; Proteins; Reaction; Reporting; Research; Resolution; Retroviridae; Role; SIV; Spumavirus; Structure; Synapses; Therapeutic; Time; Viral; Virion; Vision; Work; clinical development; clinically relevant; human model; hydroxyl group; inhibitor/antagonist; model building; novel; particle; prototype; public health relevance; structural biology; three dimensional structure; three-dimensional modeling; viral DNA; viral RNA; virology

 DESCRIPTION (provided by applicant): Integration, catalyzed by the viral integrase protein, is an essential step in the life cycle of all retroviruses, and the integrase enzyme of human immunodeficiency virus type 1 (HIV-1) is a common target of the highly active antiretroviral therapies that are used to treat AIDS patients. Integrase strand transfer inhibitors (INSTIs) have been in clinical use since 2007, and the prior iteration of this renewal application critically discovered their mechanism of action. The target of the INSTIs is the integrase-viral DNA nucleoprotein complex, also known as the intasome, and the understanding of the mechanism of therapeutic action is greatly facilitated through the study of detailed 3- dimensional structure of drug targets. Although a high-resolution structure of the HIV-1 intasome has yet to be reported, we have reported numerous structures for the prototype foamy virus (PFV) intasome, which is also inhibited by the INSTIs. Using this model, we previously described that INSTIs work by ejecting the critical deoxyadenylate residue of viral DNA and its associated 3'-hydroxyl nucleophile from the integrase active site, disarming the integration machinery. During the current funding period we extended this observation to discover that INSTIs are structural mimics of the chemical attacking and leaving groups of the DNA strand transfer reaction. Herein we provide preliminary data for a new retroviral intasome structure, and we will use this new structural information together with the structure of the PFV intasome to refine our working model of the HIV-1 intasome, the clinically relevant INSTI target. We have discovered that the most intriguing integrase drug class since the INSTIs, the allosteric integrase inhibitors (ALLINIS), inhibits HIV-1 particle maturation, implying a structural role for integrase in HIV-1 particle morphogenesis. The role of integrase in forming the infectious HIV-1 structure will be elucidated using a variety of biochemical and genetic techniques. This line of research will culminate with a biologically realistic model for the HIV-1 intasome to aid novel INSTI development and with an acute vision of the mechanism of ALLINI action, which will inform the clinical development of this new and important anti-HIV drug class.

 描述（通过应用提供）：由病毒整合蛋白催化的整合是所有逆转录病毒生命周期的重要一步，并且人类免疫缺陷病毒1型（HIV-1）的整合酶是高度活性抗逆转录病毒疗法的常见靶标，可用于治疗治疗患者。自2007年以来，整合酶链转移抑制剂（Instis）一直从事临床用途，此更新应用的事先迭代认真发现了其作用机理。该研究所的目标是综合病毒DNA核蛋白复合物，也称为核心体，对理论理论的机理的理解得到了对药物靶标的详细3维结构的研究，从而极大地支持了该理论的理论。尽管尚未报告HIV-1 Intasome的高分辨率结构，但我们报告了许多原型泡沫病毒（PFV）Intasome的结构，该结构也受到该研究所的抑制。使用该模型，我们先前描述了Instis通过从积分酶活性位点射出病毒DNA及其相关的3'-羟基核亲核核心的关键脱氧腺苷居住地，从而解除了整合机械的武装。在当前的资金期间，我们扩展了这一观察结果，发现研究所是化学攻击的结构模拟物，并留下了DNA链转移反应的组。本文中，我们为新的逆转录病毒内剂结构提供初步数据，我们将使用此新结构信息以及PFV Intasome的结构来完善我们的HIV-1 Intasome的工作模型，即临床上相关的Insti目标。我们已经发现，自该研究所（Allosteric intostric抑制剂（Allinis））以来，最有趣的整合酶药物类别抑制了HIV-1颗粒的成熟，这意味着整合酶在HIV-1颗粒形态发生中的结构作用。积分酶在形成感染性HIV-1结构中的作用将使用多种生化和遗传技术阐明。这一研究将以HIV-1 Intasome的生物学现实模型为导致，以帮助新的Insti开发以及对Allini作用机制的敏锐视野，这将为这一新的重要抗HIV药物类的临床开发提供信息。