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.

项目摘要 成熟的HIV-1衣壳核心由于其在HIV感染性中的关键作用而成为关键的抗病毒靶点 以及发现了衣壳蛋白特异性宿主限制因子，如TRIM 5 α和MX2。最近的工作已经 证明了衣壳不仅仅是简单地容纳病毒遗传物质， 复制酶它还参与并可能介导几个关键的复制事件， 包括去包被、启动逆转录、核输入、整合和逃避宿主免疫 应答对于这些过程中的每一个发生，衣壳稳定性和解离之间的微妙平衡 必须保持，证明了衣壳和病毒感染性之间的复杂联系。 成熟的衣壳晶格在蛋白酶介导的Gag多蛋白裂解后形成。基本 成熟晶格的结构元件是衣壳蛋白（CA）六聚体，其包含CA二聚体的三聚体。 富勒烯锥结构包含约250个六聚体，沿着有12个五聚体以便于闭合。突变 改变衣壳蛋白（CA）组装状态（二聚体、五聚体、六聚体或 组装的晶格）由于一个或多个复制事件的中断而导致严重的感染性缺陷。 复制也可以通过改变宿主因子结合来影响。值得注意的是，CA在以下方面的具体作用 对这些事件的理解还不很清楚，也不知道各种CA组装状态对 衣壳功能或与复制中涉及的宿主因子的相互作用。目前没有工具可用于 在体内区分CA组装状态以评估它们在复制事件期间的作用。该项目将 鉴定和表征RNA适体，所述RNA适体结合对组装的六聚体晶格特异的位点， 通过绑定到独特的溶剂暴露裂缝来区分CA组装状态， 每个独立的CA组件状态。拟议的实验利用了研究小组的 多靶适体选择的专业知识，选择后生物信息学分析的进展，细胞内 适体表达、HIV生物学和先天免疫。重要的是，拟议的方法可以广泛地 适用于对公共卫生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