Role of HIV-1 Integrase in the Late Stage of Viral Replication

HIV-1整合酶在病毒复制后期的作用

• 批准号: 8793750
• 负责人: Jacques J. Kessl
• 金额: $ 23.1万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2017-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8793750
• 关键词: Active Sites; Address; Affect; Antiviral Agents; Atomic Force Microscopy; Binding; Binding Sites; Biological; Biological Assay; Biology; Cells; Chromatin; Chromosomes; Clinical; Complex; Development; Drug resistance; Ensure; Enzymes; Exhibits; Fluorescence; Genes; Genetic; Genome; HIV-1; In Vitro; Infection; Inhibitory Concentration 50; Integrase; Integrase Inhibitors; Link; Maps; Mass Spectrum Analysis; Molecular; Molecular Sieve Chromatography; Morphology; Mutation; Nature; Nucleoproteins; Outcome Study; Patients; Phenotype; Play; Process; Protein Footprinting; Proteins; Publishing; RNA; RNA Binding; Recombinants; Reporting; Resistance; Reverse Transcription; Ribonucleoproteins; Role; Staging; Testing; Time; Viral; Virion; Virus Replication; base; clinical application; cofactor; dimer; drug discovery; electron density; improved; inhibitor/antagonist; innovation; insight; mutant; novel; public health relevance; scaffold; success; therapeutic target; transcriptional coactivator p75; viral DNA; viral RNA; virology

DESCRIPTION (provided by applicant): HIV-1 integrase (IN) is essential for viral replication and thus is an important therapeutic target. During the early stages of viral replication, a tetramer of HIV-1 IN catalyzes integration of reverse transcribed viral DNA into the host genome. The ordered multimerization of IN in the presence of viral DNA is critical for IN catalytic activity. Cellular cofactor LEDGF/p75 binds to the pre-assembled IN- viral DNA complex and tethers the nucleoprotein complex to active genes, thus ensuring effective integration. Allosteric integrase inhibitors (ALLINIs) are a novel class of integrase inhibitors that bind at the LEDGF/p75 binding site at the IN dimer interface and are capable of triggering aberrant IN multimerization. In addition, HIV-1 virions produced in the presence of ALLINIs display abnormal morphology of the virion cores and are defective for subsequent reverse transcription and integration in target cells. Interestingly, these phenotypes are similar to certain IN mutants, which have been termed class II mutants. Collectively, the studies with ALLINIs and select IN class II mutants suggest that HIV-1 IN plays a key role during late stage HIV-1 replication. However, the underlying mechanism is not clear. The present application will test the following hypothesis: ordered IN multimerization is important for its interaction with viral RNA during the late stage viral replication. Therefore, we propose the following two specific aims. Aim 1 will explore the significance of HIV-1 IN interactions with RNA for formation of the functional ribonuleoprotein complexes during the late stage viral replication. Aim 2 will characterize IN mutants that affect IN multimerization and/or IN- LEDGF/p75 binding. Mechanistic and structural details that will emerge from these studies will inform drug discovery efforts to develop improved ALLINIs for their clinical application.

描述（由申请人提供）：HIV-1整合酶（in）对于病毒复制至关重要，因此是重要的治疗靶点。在病毒复制的早期阶段，HIV-1的四聚体在催化反转录病毒DNA的催化整合到宿主基因组中。在病毒DNA存在下的有序多聚化对催化至关重要 活动。细胞辅因子LEDGF/p75与预组装的内部病毒DNA复合物结合，并将核蛋白复合物与活性基因相关，从而确保有效整合。变构积分酶抑制剂（Allinis）是一类新型的积分酶抑制剂，在IN二聚体界面处的LEDGF/p75结合位点结合，并且能够在多层化中触发异常。此外，在存在Allinis存在下产生的HIV-1病毒体显示了病毒座核的异常形态，并且在靶细胞中随后的逆转录和整合是有缺陷的。有趣的是，这些表型与已称为II类突变体的突变体中的确定性相似。总体而言，对Allinis和II类突变体中选择的研究表明，HIV-1在晚期HIV-1复制中起关键作用。但是，基本机制尚不清楚。本应用将检验以下假设：多聚化的顺序对于在晚期病毒复制中与病毒RNA的相互作用很重要。因此，我们提出以下两个具体目标。 AIM 1将探讨HIV-1与RNA相互作用在晚期病毒复制过程中形成功能性核糖蛋白复合物的重要性。 AIM 2将以影响多聚体和/或IN-LEDGF/P75结合的突变体来表征。这些研究将出现的机械和结构细节将为药物发现的努力提供依据，以改善其临床应用。