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，因此是一个重要的治疗靶点。在病毒复制的早期阶段，HIV-1 IN 四聚体催化逆转录病毒 DNA 整合到宿主基因组中。在病毒 DNA 存在的情况下，IN 的有序多聚化对于 IN 催化至关重要 活动。细胞辅因子 LEDGF/p75 与预组装的 IN-病毒 DNA 复合物结合，并将核蛋白复合物与活性基因连接，从而确保有效整合。变构整合酶抑制剂 (ALLINI) 是一类新型整合酶抑制剂，可结合 IN 二聚体界面的 LEDGF/p75 结合位点，并能够触发异常的 IN 多聚化。此外，在 ALLINI 存在的情况下产生的 HIV-1 病毒体显示出病毒体核心的异常形态，并且在随后的逆转录和在靶细胞中的整合方面存在缺陷。有趣的是，这些表型与某些 IN 突变体相似，被称为 II 类突变体。总的来说，ALLINI 和选择的 IN II 类突变体的研究表明，HIV-1 IN 在 HIV-1 复制的晚期阶段发挥着关键作用。然而，其根本机制尚不清楚。本申请将检验以下假设：有序的IN多聚化对于其在病毒复制后期与病毒RNA的相互作用非常重要。因此，我们提出以下两个具体目标。目标 1 将探讨 HIV-1 IN 与 RNA 相互作用对于病毒复制后期功能性核糖核蛋白复合物形成的重要性。目标 2 将表征影响 IN 多聚化和/或 IN-LEDGF/p75 结合的 IN 突变体。这些研究中出现的机制和结构细节将为药物发现工作提供信息，以开发改进的 ALLINI 用于临床应用。

项目成果

Indole-based allosteric inhibitors of HIV-1 integrase.

基于吲哚的 HIV-1 整合酶变构抑制剂。

• 发表时间: 2016
• 影响因子: 2.7
• 作者: Patel, Pratiq A;Kvaratskhelia, Nina;Mansour, Yara;Antwi, Janet;Feng, Lei;Koneru, Pratibha;Kobe, Mathew J;Jena, Nivedita;Shi, Guqin;Mohamed, Mosaad S;Li, Chenglong;Kessl, Jacques J;Fuchs, James R
• 通讯作者: Fuchs, James R

Methods for the Analyses of Inhibitor-Induced Aberrant Multimerization of HIV-1 Integrase.

抑制剂诱导的 HIV-1 整合酶异常多聚化的分析方法。

• 发表时间: 2016
• 作者: Kessl, Jacques J;Sharma, Amit;Kvaratskhelia, Mamuka
• 通讯作者: Kvaratskhelia, Mamuka