Retroviral capsid recognition by TRIM5alpha restriction factors

TRIM5alpha 限制因子识别逆转录病毒衣壳

• 批准号: 8732420
• 负责人: DMITRI N IVANOV
• 金额: $ 33.12万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 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 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8732420
• 关键词: Accounting; Acquired Immunodeficiency Syndrome; Affect; Affinity; Amino Acids; Anti-Retroviral Agents; Binding; Biological Assay; C-terminal; Capsid; Capsid Proteins; Cells; Charge; Chemicals; Complex; Computing Methodologies; Defect; Docking; Epitopes; Evolution; Genetic Materials; Genome; Goals; HIV; Human; Human Activities; Infection; Intervention; Knowledge; Light; Macaca mulatta; Maps; Measurement; Measures; Mediating; Modeling; Molecular; Mutagenesis; Mutation; NMR Spectroscopy; Natural Immunity; Organism; Pan Genus; Prevention; Primate Lentiviruses; Primates; Property; Proteins; Race; Refractory; Research; Research Proposals; Resolution; Retroviridae; Reverse Transcription; SIV; Specificity; Spectrum Analysis; Structure; Surface; TRIM Gene; Testing; Titrations; Tropism; Ursidae Family; Variant; Viral; X-Ray Crystallography; arm; base; biophysical techniques; computerized tools; insight; molecular dynamics; novel; pandemic disease; pathogen; public health relevance; restraint; retroviral-mediated; success; virus genetics

DESCRIPTION: TRIM?? restriction factors bind retroviral capsids after cell entry and restrict retroviral infection by blocking reverse transcription and/or integration of the viral genetic material. The C-terminal SPRY domain of TRIM?? is believed to form most of the capsid-TRIM?? interface as species-specific sequence variations within the SPRY domain account for differences in the viral specificity of the TRIM??-mediated restriction. Most notably, recent evolution of the human TRIM?? SPRY resulted in the variant that has poor affinity for the HIV capsid, the vulnerability that contributed to the emergence of the AIDS pandemic when the simian immunodeficiency virus (SIV) passed from chimpanzees into a human host. Mutation or deletion of a single amino acid residue (R332) within the SPRY domain restores HIV restriction by the human TRIM?? variant. TRIM?? SPRY domains have proven refractory to structural and biophysical studies limiting our understanding of retroviral capsid recognition by TRIM??. This research proposal seeks to fill that knowledge gap by solving the structures of several primate TRIM?? SPRY variants with distinct retroviral specificities and characterizing their interactions with the cognate retroviral capsids. The goal of the proposed research is to understand the mechanism of capsid recognition by TRIM?? restriction factors, elucidate why the human TRIM?? variant is not potent against HIV and to explore whether this defect could be alleviated by pharmacological or other means. The following three specific aims will be pursued: (1) The structures of the rhesus, human and possibly other TRIM?? SPRY domains will be solved using a novel experimental approach that combines NMR spectroscopy and X-ray crystallography. (2) Interaction of several SPRY-capsid pairs will be characterized using a variety of biophysical approaches. (3) High-resolution models of the SPRY-capsid complexes will be generated using computational methods and tested using mutagenesis and restriction assays.

描述：修剪??限制因子通过阻断病毒遗传材料的逆转录和/或整合，结合细胞进入后逆转录病毒衣壳并限制逆转录病毒感染。修剪的C末端SPRY域？据信会形成大部分的衣壳三体？界面作为SPRY域内的物种特异性序列变化，解释了内饰介导的限制的病毒特异性的差异。最值得注意的是，人类修剪的最新演变？ Spry导致了对HIV Capsid的亲和力差的变体，当Simian免疫缺陷病毒（SIV）从黑猩猩传递到人类宿主时，导致艾滋病大流行的脆弱性。 Spry结构域中单个氨基酸残基（R332）的突变或缺失可通过人体修剪恢复HIV限制？变体。修剪？？ Spry结构域已证明对结构和生物物理研究的难治性限制了我们通过修剪的逆转录病毒衣壳识别的理解。这项研究建议试图通过解决几个灵长类动物装饰的结构来填补这一知识差距？具有不同逆转录病毒特异性的Spry变体，并表征了它们与同源逆转录病毒衣壳的相互作用。拟议的研究的目的是了解修剪的Capsid识别机制？限制因素，阐明为什么人类修剪？变体对艾滋病毒没有效力，并探索该缺陷是否可以通过药理或其他手段来缓解。将追求以下三个特定目标：（1）恒河所，人类以及可能的其他装饰的结构？ Spry结构域将使用新型的实验方法来解决，该方法结合了NMR光谱和X射线晶体学。 （2）使用多种生物物理方法的几个Spry-Capsid对的相互作用将被表征。 （3）将使用计算方法生成Spry-Capsid复合物的高分辨率模型，并使用诱变和限制测定进行测试。