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.

摘要 HIV-1整合酶（IN）是病毒复制所必需的，并催化病毒DNA整合到细胞内。 寄主染色体最近报道了一类新的变构IN抑制剂（ALLINI）， 能够通过促进异常IN多聚化来抑制酶功能。没想到在 感染的细胞，这些化合物通过触发HIV-1复制的晚期， 显著的病毒核心形态缺陷。这种表型表明，除了已知的 在HIV-1复制的早期阶段，IN也起着重要的非催化作用， 在病毒复制的后期阶段发挥作用。我们的初步研究表明，IN与 在体外和适当成熟的病毒体中选择高度结构化的病毒RNA元件。此外，本发明还提供了一种方法， 我们能够观察到ALLINI能够调节病毒体中的这种相互作用。的 以下两个目标将扩展这些初步意见。目标1将绘制病毒RNA结合图 IN上的位点，并确定所鉴定的相互作用氨基酸的功能意义。目标2将 确定ALLINI在病毒体形态发生期间如何影响IN与病毒RNA的相互作用。采取 我们将共同使用病毒学和生物化学方法的组合，使我们能够检查 IN与病毒RNA基因组的相互作用和适当成熟的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IN如何的理解。 在HIV-1复制的后期阶段的功能，以及阐明的作用方式， 最近发现了阿利尼斯。