Structural and Functional Integration of TRIM5alpha Domains for HIV Capsid Binding

用于 HIV 衣壳结合的 TRIM5alpha 结构域的结构和功能整合

• 批准号: 9247716
• 负责人: Jonathan Mark Wagner
• 金额: $ 6.1万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9247716
• 关键词: Amino Acid Motifs; Attention; Behavior; Binding; Binding Sites; Capsid; Capsid Proteins; Cells; Chromosomes, Human, Pair 11; Coiled-Coil Domain; Complement; Complex; Crystallization; Data; Data Set; Defense Mechanisms; Dimerization; Disulfides; Electron Spin Resonance Spectroscopy; Electrons; Epitopes; Equilibrium; Family member; Fluorescence Resonance Energy Transfer; Goals; HIV; HIV Infections; Homodimerization; Homologous Gene; Imagery; Immune system; In Vitro; Income; Individual; Intercept; Invaded; Investigation; Label; Link; Location; Measures; Mediating; Modeling; Molecular; Molecular Conformation; Molecular Structure; Movement; Nucleosome Core Particle; Pattern recognition receptor; Positioning Attribute; Protein Family; Proteins; Recombinants; Research; Resolution; Rest; Reverse Transcription; Role; Signal Pathway; Site; Spin Labels; Structure; TRIM Motif; TRIM5 gene; Testing; Therapeutic; Time; Tube; Viral; Virus; Virus Replication; design; dimer; experimental study; flexibility; improved; information model; monomer; network models; particle; premature; prevent; protein structure; public health relevance; two-dimensional

 DESCRIPTION (provided by applicant): The Tripartite Motif (TRIM) proteins are important components of first line cellular defense mechanisms against invading viruses. One TRIM family member, TRIM5α, directly binds and disassembles incoming human immunodeficiency virus (HIV) cores, preventing reverse transcription and inhibiting viral replication. At the same time, TRIM5α acts as a pattern recognition receptor that stimulates downstream proinflammatory signaling pathways. In order to accomplish these, TRIM5α first dimerizes through its coiled-coil domain and the dimers further assemble via their B-box 2 domains into a two-dimensional hexagonal lattice that surrounds HIV core particles. The hexagonal lattice contains regularly spaced capsid binding SPRY domains that mediate direct interactions with the proteinaceous HIV capsid shell. Each individual SPRY domain interacts very weakly, but the assembled TRIM network binds avidly by aligning SPRY domains with regularly spaced capsid epitopes. Recent structures of individual TRIM5α domains have suggested that the spacing and orientation of SPRY domains in TRIM5α might be controlled by a four-way packing interaction between two SPRY domains resting on two coiled-coil domains. However, a structure of the TRIM5α dimer including both coiled-coil and SPRY domains is necessary to confirm this. Furthermore, it appears that SPRY domains may be able to release from the coiled-coil domains with the intervening linker 2 regions acting as a flexible tether. This study will investigate the structureof the TRIM5α coiled-coil and SPRY dimer and the dynamic movements that may be associated with HIV capsid binding. We expect to discover the locations and orientations of capsid binding SPRY domains within the network surrounding HIV capsid particles. This will inform our understanding of the molecular defenses used by the cell to destroy viruses and may ultimately improve our ability to design therapeutics that mimic or complement these complex cellular machines.

 描述(申请人提供)：三方基序(TRIM)蛋白是针对入侵病毒的一线细胞防御机制的重要组成部分。TRIM家族成员之一，TRIM5α，直接结合和分解传入的人类免疫缺陷病毒核心，防止反转录和抑制病毒复制。同时，TRIM5α作为一种模式识别受体刺激下游的促炎信号通路。为了完成这些，TRIM5HIV首先通过其盘绕的线圈结构域进行二聚，二聚体通过它们的B-box 2结构域进一步组装成围绕α核心颗粒的二维六角形晶格。六角形晶格包含规则间隔的衣壳结合SPRY结构域，这些结构域介导与蛋白质类HIV衣壳的直接相互作用。每个单独的spry结构域相互作用非常弱，但组装的TRIM网络通过将spry结构域与规则间隔的衣壳表位比对而强烈结合。最近单个TRIM5α结构域的结构表明，TRIM5α中SPRY结构域的间距和取向可能由两个螺旋线圈结构域上的两个SPRY结构域之间的四向堆积作用控制。然而，为了证实这一点，TRIM5α二聚体的结构同时包括螺旋结构域和SPRY结构域是必要的。此外，看起来SPRY结构域可能能够从盘绕的线圈结构域中释放，中间的连接体2区域起到柔性系绳的作用。本研究将研究TRIM5二聚体和α卷曲螺旋的结构，以及可能与HIV衣壳结合有关的动态运动。我们希望发现HIV衣壳颗粒周围网络中衣壳结合spry结构域的位置和方向。这将使我们了解细胞用来摧毁病毒的分子防御，并最终可能提高我们设计模仿或补充这些复杂细胞机器的疗法的能力。