Role of the B-box-v-1 Restriction by TRIM5alpha proteins

TRIM5alpha 蛋白对 B-box-v-1 限制的作用

• 批准号: 8015748
• 负责人: Felipe Diaz-Griffero
• 金额: $ 24.9万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8015748
• 关键词: Address; Affect; Award; Binding; Biological Assay; Boxing; Capsid; Capsid Proteins; Cell Nucleus; Cells; Coiled-Coil Domain; Cytoplasm; Cytosol; Dana-Farber Cancer Institute; Environment; Event; Gene Expression; Genome; Genomics; Goals; HIV; HIV-1; Homologous Gene; Human; Human Activities; Immunologic Deficiency Syndromes; Infection; Integration Host Factors; Laboratories; Macaca mulatta; Mediating; Mentors; Molecular; Mutagenesis; Mutation; Phase; Primates; Process; Production; Proteins; RNA Splicing; Research; Research Personnel; Role; Specificity; Structure; TRIM Motif; Testing; Time; Training; Variant; Viral; Viral Genes; improved; monomer; mutant; novel therapeutics; particle; research study; viral RNA

The recently discovferecl rhesus monkey protein;TRIM5< is responsible for the strong restriction imposed to human immunodeficiency vims (HIV-1) infection by these primates. We have con-elated the restriction of HIV-1 by TRiM5<rhwith a deei^ease of viral core stability in the cytoplasm during infection. This suggested that TRll^5<(1ipNteinsiblock;retroviral replication bychahging the stability of the incoming retroviral core into the host cell. Separately,! changes in the stability of the incoming retroviral core achieved by HIV-1 capsid mutagenesis resulted also in a loss of ihfectivity; these findings suggested that modulation of viral core The recently discovered rhesus monkey protein TRIMSalpha (TRIM5alpha-rh) is responsible for the strong restriction imposed to human immunodeficiency virus (HIV-1) infection by these primates. We have correlated the restriction of HIV-1 by TRIM5alpha-rh with a decrease of viral core stability in the cytoplasm during infection. This suggested that TRIM5alpha-rh proteins block retroviral replication by changing the stability of the incoming retroviral core into the host cell. Separately, changes in the stability of the incoming retroviral core achieved by HIV-1 capsid mutagenesis results also in a loss of infectivity. Collectively, these findings suggested that modulation of viral core stability is detrimental for infection. Remarkably, we have found mutations in the B-box 2 domain of TRIM5alpha-rh that bind the HIV-1 incoming viral core, yet the stability or infectivity of the core is not affected. This particular set of B-box 2 domain mutants could bind capsid and oligomerize when compared to wild type. For these reasons, we hypothesize that binding to capsid by these mutants might be achieved differently than wild-type. In this proposal, we will test the hypothesis that cooperative binding of TRIM5alpha-rh trimers to the retroviral core results in core destabilization and inhibition of infection; and that cooperative binding of TRIM5alpha-rh trimers to the viral core is mediated by B-box 2-B-box 2 inter-trimer interactions. To investigate this hypothesis we will: 1) identify the B-box 2 determinants for retroviral restriction by structure-function studies; 2) elucidate the role of B-box 2-B-box 2 interactions in retroviral restriction by TRIM5alpha-rh; and 3) assay cooperative binding of TRIM5alpha-rh trimers to the HIV-1 capsid. The completion of the proposed research will help me to achieve my long-term goals of becoming an independent investigator in the field of HIV-1/AIDS. For this reason the Dana-Farber Cancer Institute in combination with the laboratory of Dr. Joseph Sodroski is the adequate environment to pursue this goal. In addition, several training classes will be taken in order to smooth the transition from the mentored phase to the independent phase of the award.

最近发现的恒河猴蛋白；TRIM5< 是造成对 这些灵长类动物感染了人类免疫缺陷病毒（HIV-1）。我们已将限制 TRiM5的HIV-1感染期间细胞质中的病毒核心稳定性降低。这建议 TRll^5<(1ipNteinsiblock;通过改变传入逆转录病毒核心的稳定性来进行逆转录病毒复制 宿主细胞。分别地，！ HIV-1衣壳实现的传入逆转录病毒核心稳定性的变化 诱变也会导致感染性丧失；这些发现表明病毒核心的调节 最近发现的恒河猴蛋白 TRIMSalpha (TRIM5alpha-rh) 负责强 对这些灵长类动物感染人类免疫缺陷病毒（HIV-1）的限制。我们有 TRIM5alpha-rh 对 HIV-1 的限制与细胞质中病毒核心稳定性的降低相关 感染期间。这表明 TRIM5alpha-rh 蛋白通过改变 进入宿主细胞的逆转录病毒核心的稳定性。另外，输入稳定性的变化 HIV-1衣壳诱变实现的逆转录病毒核心也会导致感染性丧失。总的来说，这些 研究结果表明，病毒核心稳定性的调节对感染有害。值得注意的是，我们有 发现 TRIM5alpha-rh 的 B-box 2 结构域发生突变，该突变与 HIV-1 传入病毒核心结合，但 核心的稳定性或感染性不受影响。这组特殊的 B-box 2 结构域突变体可以结合 与野生型相比，衣壳和寡聚化。由于这些原因，我们假设结合 这些突变体的衣壳可能与野生型不同。在本提案中，我们将测试 假设 TRIM5alpha-rh 三聚体与逆转录病毒核心的协同结合导致核心 不稳定和感染抑制； TRIM5alpha-rh 三聚体与病毒的协同结合 核心由 B-box 2-B-box 2 三聚体间相互作用介导。为了研究这个假设，我们将：1) 通过结构-功能研究确定逆转录病毒限制的 B-box 2 决定因素； 2）阐明角色 TRIM5alpha-rh 逆转录病毒限制中的 B-box 2-B-box 2 相互作用； 3) 测定协同结合 HIV-1 衣壳的 TRIM5alpha-rh 三聚体。完成拟议的研究将帮助我实现 我的长期目标是成为 HIV-1/AIDS 领域的独立研究者。为此 丹纳法伯癌症研究所与 Joseph Sodroski 博士的实验室相结合是足够的 追求这一目标的环境。此外，还将举办多门培训课程，以确保顺利进行 奖项从指导阶段过渡到独立阶段。