Mode of Action of Allosteric HIV-1 Integrase Inhibitors during Late Steps of Viral Replication

变构 HIV-1 整合酶抑制剂在病毒复制后期的作用模式

• 批准号: 9204097
• 负责人: Jacques J. Kessl
• 金额: $ 18.44万
• 依托单位: UNIVERSITY OF SOUTHERN MISSISSIPPI
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-04 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9204097
• 关键词: Affect; Affinity; Amino Acids; Anti-Retroviral Agents; Antiviral Agents; Atomic Force Microscopy; Binding; Binding Sites; Biochemical; Biology; Capsid; Cells; Chromosomes; Clinical; Complex; Defect; Development; Electrons; Elements; Enzymes; Foundations; Future; Genome; HIV; HIV-1; HIV-1 integrase; In Vitro; Infection; Integrase; Integrase Inhibitors; Link; Maps; Mass Spectrum Analysis; Membrane; Morphogenesis; Morphology; Nucleosome Core Particle; Patients; Phenotype; Physical condensation; Play; Protein Footprinting; Publishing; RNA Binding; Recombinants; Reporting; Resistance; Ribonucleoproteins; Role; Staging; Structure; Time; Viral; Virion; Virus; Virus Replication; abstracting; base; cofactor; improved; inhibitor/antagonist; mutant; novel; particle; pyridine; quinoline; research study; success; therapeutic target; transcriptional coactivator p75; viral DNA; viral RNA; virology

Abstract HIV-1 integrase (IN) is essential for viral replication and catalyzes integration of viral DNA into the host chromosome. A new class of allosteric IN inhibitors (ALLINIs) has recently been reported to be capable of inhibiting the enzyme function by promoting aberrant IN multimerization. Unexpectedly, in infected cells, these compounds potently impaired the late stage of HIV-1 replication by triggering significant viral core morphology defects. This phenotype suggested that in addition to its known catalytic function during the early steps of HIV-1 replication, IN also plays an important, non-catalytic role during the late steps of viral replication. Our preliminary studies reveal that IN tightly binds to select highly structured viral RNA elements both in vitro and in properly matured virions. In addition, we were able to observe that an ALLINI is capable of modulating this interaction in the virion. The following two aims will extend these preliminary observations. Aim 1 will map the viral RNA binding sites on IN and determine the functional significance of the identified interacting amino acids. Aim 2 will determine how ALLINIs affect IN interactions with viral RNA during virion morphogenesis. Taken together, we will use a combination of virology and biochemical approaches to allow us to examine the potential link between IN interactions with viral RNA genome and formation of properly matured virions. Structural details that will emerge from these studies will increase our understanding of how IN functions during the late stage of HIV-1 replication as well as elucidate the mode of action for the recently discovered ALLINIs.

抽象的 HIV-1整合酶（IN）对于病毒复制和催化病毒DNA的整合至关重要 宿主染色体。据报道 能够通过促进多种化的异常来抑制酶功能。出乎意料的是 感染细胞，这些化合物通过触发HIV-1复制的后期有效地损害了 明显的病毒核心形态缺陷。这种表型表明，除了其已知 在HIV-1复制的早期步骤中的催化功能，也起着重要的，非催化的作用 在病毒复制的后期步骤中的作用。我们的初步研究表明，紧密结合 在体外和正确成熟的病毒体中选择高度结构化的病毒RNA元素。此外， 我们能够观察到Allini能够调节病毒体中的这种相互作用。这 以下两个目标将扩展这些初步观察。 AIM 1将绘制病毒RNA结合 位置上并确定已鉴定的相互作用氨基酸的功能意义。 AIM 2意志 确定在病毒形态发生过程中与病毒RNA相互作用的相互作用中的Allinis如何影响。拍摄 我们将共同使用病毒学和生化方法的结合，使我们能够检查 与病毒RNA基因组相互作用与正确成熟病毒体的形成之间的潜在联系。 从这些研究中出现的结构细节将增加我们对如何中的理解 在HIV-1复制后期的功能，并阐明 最近发现了Allinis。