RNA Aptamers that Differentiate Among HIV-1 Capsid Assembly States

区分 HIV-1 衣壳组装状态的 RNA 适体

• 批准号: 9303240
• 负责人: Donald H Burke
• 金额: $ 23.03万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-24 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9303240
• 关键词: Antiviral Agents; Binding; Binding Sites; Bioinformatics; Biological; Biology; Capsid; Capsid Proteins; Cells; Cone; Defect; Dimerization; Dissociation; Elements; Enzymes; Equilibrium; Event; Fullerenes; Future; Genetic Materials; HIV; HIV-1; Health; Immune response; In Vitro; Integration Host Factors; Investigation; Lead; Life Cycle Stages; Link; Mediating; Methods; Mutation; Natural Immunity; Nuclear Import; Peptide Hydrolases; Pharmaceutical Preparations; Process; Proteins; Public Health; RNA; Research; Reverse Transcription; Role; Site; Solvents; Structure; Surface; Viral; Viral Pathogenesis; Virus; Virus Replication; Work; aptamer; dimer; experimental study; gag Gene Products; in vivo; small molecule; structural biology; tool; virus genetics

Project Summary The mature HIV-1 capsid core has emerged as a key antiviral target because of its critical role in HIV infectivity and the discovery of capsid-specific host restriction factors, such as TRIM5α and MX2. Recent work has demonstrated that capsid does much more than simply house the viral genetic material and required replication enzymes. It also participates in and may mediate several critical replication events, including uncoating, initiation of reverse transcription, nuclear import, integration, and evasion of host immune responses. For each of these processes to occur, a delicate balance between capsid stability and dissociation must be maintained, demonstrating an intricate link between capsid and viral infectivity. The mature capsid lattice is formed following protease-mediated cleavage of the Gag polyprotein. The basic structural element of the mature lattice is a capsid protein (CA) hexamer, comprising a trimer of CA dimers. The fullerene cone structure contains ~250 hexamers, along with 12 pentamers to facilitate closing. Mutations that alter the relative stability of capsid protein (CA) assembly states (dimers, pentamers, hexamers, or the assembled lattice) result in severe infectivity defects due to disruption of one or more replication events. Replication can also be impacted through alteration of host factor binding. Notably, the specific roles of CA in these events is not well understood, and it is unknown how the various CA assembly states contribute to capsid function or interactions with host factors involved in replication. There are currently no tools available to differentiate CA assembly states in vivo to assess their role during replication events. This project will identify and characterize RNA aptamers that bind sites specific to the assembled hexamer lattice and differentiate among CA assembly states by binding to unique solvent-exposed crevices that define each independent CA assembly state. The proposed experiments capitalize on the research team's expertise in poly-target aptamer selection, advances in post-selection bioinformatics analysis, intracellular aptamer expression, HIV biology, and innate immunity. Importantly, the proposed approach could be widely applicable to other viruses of importance to public health.

项目概要 成熟的 HIV-1 衣壳核心因其在 HIV 感染中的关键作用而成为关键的抗病毒靶点 衣壳特异性宿主限制因子的发现，例如 TRIM5α 和 MX2。 衣壳的作用远不只是简单地展示病毒的遗传物质和所需的 它也参与并可能介导一些关键的复制事件， 包括脱壳、逆转录启动、核输入、整合和逃避宿主免疫 对于每个过程的发生，衣壳稳定性和解离之间存在微妙的平衡。 必须维持，证明衣壳和病毒感染性之间存在复杂的联系。 成熟的衣壳晶格是在蛋白酶介导的 Gag 多蛋白裂解后形成的。 成熟晶格的结构元件是衣壳蛋白（CA）六聚体，包含CA二聚体的三聚体。 富勒烯锥体结构包含约 250 个六聚体，以及 12 个五聚体以促进突变。 改变衣壳蛋白 (CA) 组装状态（二聚体、五聚体、六聚体或 组装晶格）由于一个或多个复制事件的破坏而导致严重的感染性缺陷。 宿主因子结合的改变也会影响复制，值得注意的是，CA 在其中的具体作用。 这些事件还没有被很好地理解，并且不知道各个 CA 议会州如何做出贡献 衣壳功能或与复制相关的宿主因子的相互作用目前没有可用的工具。 该项目将区分体内 CA 组装状态以评估它们在复制事件中的作用。 识别和表征与组装的六聚体晶格特异位点结合的 RNA 适体 通过结合到独特的暴露于溶剂的缝隙来区分 CA 组装状态 每个独立的 CA 组装状态都利用了研究小组的实验。 多目标适体选择方面的专业知识、选择后生物信息学分析的进展、细胞内 重要的是，所提出的方法可以广泛应用。 适用于对公共卫生具有重要意义的其他病毒。

项目成果

GS-CA Compounds: First-In-Class HIV-1 Capsid Inhibitors Covering Multiple Grounds.

GS-CA 化合物：涵盖多种领域的一流 HIV-1 衣壳抑制剂。

• DOI: 10.3389/fmicb.2019.01227
• 发表时间: 2019
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Singh,Kamal;Gallazzi,Fabio;Hill,KyleJ;Burke,DonaldH;Lange,MargaretJ;Quinn,ThomasP;Neogi,Ujjwal;Sönnerborg,Anders
• 通讯作者: Sönnerborg,Anders