Structural Virology of Tripartite Motif Proteins

三联基序蛋白的结构病毒学

• 批准号: 10888752
• 负责人: Owen Pornillos
• 金额: $ 58.8万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-15 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10888752
• 关键词: Acceleration; Affect; Amino Acid Motifs; Avidity; Binding; Binding Proteins; Biochemical; Biology; Capsid; Cell Nucleus; Cell membrane; Cell-Free System; Cells; Collaborations; Competence; Complex; Cryoelectron Microscopy; Dependence; Development; Dissociation; Electrons; Engineering; Event; Funding; Goals; HIV; HIV-1; HIV/AIDS; Human; In Vitro; Infection; Intercept; Interferons; Knowledge; Learning; Link; Maps; Measurement; Mediating; Modeling; Molecular; Mutation; Natural Immunity; Pathway interactions; Pattern; Pattern Recognition; Pattern recognition receptor; Pharmaceutical Preparations; Phenocopy; Phenotype; Proteins; Recombinants; Reporting; Resolution; Retroviridae; Reverse Transcription; Role; Signal Transduction; Site; Specificity; Structure; System; TRIM Motif; TRIM5 gene; Techniques; Testing; Travel; Ubiquitin; Ubiquitination; Viral; Virus; Visualization; Work; X-Ray Crystallography; cross-species transmission; dimer; fighting; human pathogen; inhibitor; insight; integration site; negative affect; novel; paralogous gene; pharmacologic; premature; reconstitution; ubiquitin-protein ligase; virology

PROJECT SUMMARY Tripartite motif (TRIM) proteins are ubiquitin E3 ligases that function in development and innate immunity. TRIM5a and TRIMCyp proteins (collectively termed TRIM5) recognize and deactivate the incoming cores of retroviruses such as HIV- 1. Indeed, human TRIM5a is an important component of interferon-mediated control of HIV-1 infection. Previous work from our group suggested an unprecedented lattice-to-lattice mode of pattern recognition, in which TRIM5a dimers use the viral capsid surrounding the viral core as a template for higher-order assembly, ultimately forming a cage around the core. In the current funding period of this R01 project, we solved the first structure of the TRIM5a/capsid superlattice, developed and optimized a recombinant assembly and ubiquitination system for TRIM5/capsid complexes, and collaborated in studies that reconstituted the early events of HIV-1 replication in a cell-free system. In this renewal application, we now propose to: Aim 1, extend the resolution of our TRIM5a/capsid superlattice structure, in order to visualize contacts between the arrayed TRIM5a protein dimers and their bound viral CA protein hexamer subunits. Aim 2, utilize our novel in vitro ubiquitination and HIV-1 replication systems as experimental platforms to elucidate how TRIM5 deactivates its caged viral core. Aim 3, understand a novel mechanism of TRIM5 recognition, in which the homologous TRIM34 protein directs human TRIM5a to bind and inactive HIV-1 capsids bearing the N74D mutation. Collectively, these studies will fill in important missing gaps in our understanding of how TRIM5 proteins inhibit HIV-1 replication. Our developing understanding of TRIM5 has been giving important insights on HIV-1 capsid vulnerabilities and efforts to develop drugs that exploit these vulnerabilities.

项目摘要 三方基序（TRIM）蛋白是在发育和先天免疫中起作用的泛素E3连接酶。 trim5a和 三膜蛋白（统称为TRIM5）识别并停用逆转录病毒的核心（例如HIV） 1。的确，人trim5a是干扰素介导的HIV-1感染控制的重要组成部分。以前的工作 我们小组提出了前所未有的晶格 - 晶格模式的模式识别模式，其中Trim5a二聚体使用 围绕病毒芯作为高阶组件的模板的病毒式衣壳，最终在周围形成一个笼子 核。在此R01项目的当前资金期间，我们解决了TRIM5A/CAPSID超晶格的第一个结构， 开发并优化了用于Trim5/capsid复合物的重组组装和泛素化系统，以及 在无单元系统中重新建立了HIV-1复制的早期事件的研究中进行了合作。在此续约中 应用程序，我们现在建议：AIM 1，扩展trim5a/capsid超晶格结构的分辨率，以便为了 可视化阵列的TRIM5A蛋白二聚体及其结合的病毒CA蛋白己酰基亚基之间的接触。目的 2，利用我们的新型体外泛素化和HIV-1复制系统作为实验平台，以阐明如何 TRIM5停用其笼子病毒核心。 AIM 3，了解TRIM5识别的新型机制，其中 同源TRIM34蛋白指导人trim5a结合和无活跃的HIV-1衣壳带有N74D突变。 总的来说，这些研究将填补我们对TRIM5蛋白如何抑制HIV-1的重要缺失空白 复制。我们对TRIM5的不断发展一直在对HIV-1 CAPSID脆弱性提供重要的见解 以及开发利用这些漏洞的药物的努力。