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（申请人提供）：由病毒整合酶蛋白催化的整合是所有逆转录病毒生命周期中必不可少的步骤，而人类免疫缺陷病毒1型（HIV-1）的整合酶是高度整合的共同目标。用于治疗艾滋病患者的主动抗逆转录病毒疗法自 2007 年以来一直在临床使用，并且该疗法的前一版本。更新应用批判性地发现了 INSTI 的作用靶点是整合酶-病毒 DNA 核蛋白复合物，也称为整合体，通过详细的 3 维结构的研究极大地促进了对治疗作用机制的理解。虽然 HIV-1 嵌体的高分辨率结构尚未报道，但我们已经报道了原型泡沫病毒 (PFV) 嵌体的许多结构，它也受到 HIV-1 嵌体的抑制。使用该模型，我们之前描述了 INSTI 通过从整合酶活性位点排出病毒 DNA 的关键脱氧腺苷酸残基及其相关的 3'-羟基亲核体，解除整合机制。发现 INSTI 是 DNA 链转移反应的化学攻击和离去基团的结构模拟。在此，我们为新的逆转录病毒嵌体结构提供了初步数据。使用这一新的结构信息和 PFV 整合酶的结构来完善我们的 HIV-1 整合酶工作模型，这是临床相关的 INSTI 靶标。我们发现了自 INSTI 以来最有趣的整合酶药物类别，即变构整合酶抑制剂（ ALLINIS），抑制 HIV-1 颗粒成熟，暗示整合酶在 HIV-1 颗粒形态发生中的结构作用 整合酶在形成感染性 HIV-1 中的作用。该系列研究将通过多种生化和遗传技术来阐明 HIV-1 嵌体的生物学真实模型，以帮助新型 INSTI 的开发，并对 ALLINI 作用机制进行敏锐的洞察，从而为人们提供信息。这一新型重要的抗艾滋病毒药物的临床开发。