Retroviral capsid recognition by TRIM5alpha restriction factors

TRIM5alpha 限制因子识别逆转录病毒衣壳

基本信息

批准号：
9262531
负责人：
DMITRI N IVANOV
金额：
$ 4.18万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
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/9262531
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 Health 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 Testing Titrations Tropism Ursidae Family Variant Viral X-Ray Crystallography arm base biophysical analysis biophysical techniques computerized tools insight molecular dynamics novel pandemic disease pathogen restraint retroviral-mediated success virus genetics

项目摘要

DESCRIPTION: TRIM5α 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 TRIM5α is believed to form most of the capsid-TRIM5α interface as species-specific sequence variations within the SPRY domain account for differences in the viral specificity of the TRIM5α-mediated restriction. Most notably, recent evolution of the human TRIM5α 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 TRIM5α variant. TRIM5α SPRY domains have proven refractory to structural and biophysical studies limiting our understanding of retroviral capsid recognition by TRIM5α. This research proposal seeks to fill that knowledge gap by solving the structures of several primate TRIM5α 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 TRIM5α restriction factors, elucidate why the human TRIM5α 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 TRIM5α 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：TRIM5α限制因子通过阻断病毒遗传材料的逆转录和/或整合来结合细胞进入后逆转录病毒衣壳并限制逆转录病毒感染。据信，TRIM5α的C末端SPRY结构域形成了大多数Capsid-Trim5α界面，作为Spry结构域中规格特异性序列变化，认为TRIM5α介导的限制的病毒特异性差异。最值得注意的是，人类TRIM5αSpry的最新演变导致这种变体对HIV Capsid的亲和力较差，当Simian免疫缺陷病毒（SIV）从黑猩猩传入人类宿主时，艾滋病大流行的脆弱性导致了艾滋病大流行的出现。人体TRIM5α变体在Spry结构域恢复内的单个氨基酸住宅（R332）的突变或缺失。 TRIM5αSPRY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对TRIM5α对逆转录病毒衣壳识别的理解。这项研究建议旨在通过解决具有不同逆转录病毒特异性的几种主要TRIM5αSPRY变体的结构来填补该知识差距，并表征其与同源逆转录病毒胶囊的相互作用。拟议的研究的目的是了解TRIM5α限制因素识别衣壳的机制，阐明为什么人类TRIM5α变体没有潜在的抗HIV，并探讨该缺陷是否可以通过药物或其他手段来缓解。将采用以下三个特定目标：（1）将使用新型的实验方法来解决恒河类，人类和可能的其他TRIM5αSpry结构域的结构，该方法结合了NMR光谱和X射线晶体学。（2）将使用计算方法生成几个Spry-Capsid对的相互作用（3）Spry-Capsid复合物的高分辨率模型，并使用诱变和限制分析进行了测试。