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

最近发现的恒河猴蛋白质TRIM 5 <负责对 人类免疫缺陷病毒（HIV-1）感染。我们已经联系到了 在感染期间，TRIM 5 <rhHIV-1在细胞质中的病毒核心稳定性降低。这表明 通过改变进入的逆转录病毒核心的稳定性， 宿主细胞分开，！通过HIV-1衣壳实现的传入逆转录病毒核心稳定性的变化 诱变也导致了免疫力的丧失;这些发现表明病毒核心的调节 最近发现的恒河猴蛋白TRIMSalpha（TRIM 5alpha-rh）是负责强 这些灵长类动物对人类免疫缺陷病毒（HIV-1）感染的限制。我们有 TRIM 5 α-rh对HIV-1的限制与细胞质中病毒核心稳定性的降低相关 在感染期间。这表明TRIM 5alpha-rh蛋白通过改变逆转录病毒的复制来阻止逆转录病毒的复制 进入宿主细胞的逆转录病毒核心的稳定性。另外，传入的稳定性的变化 通过HIV-1衣壳诱变获得的逆转录病毒核心也导致感染性丧失。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是有害的。值得注意的是， 在TRIM 5 α-rh的B-box 2结构域中发现了结合HIV-1传入病毒核心的突变， 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，我们假设绑定到 通过这些突变体的衣壳可能与野生型不同地实现。在本提案中，我们将测试 假设TRIM 5 α-rh三聚体与逆转录病毒核心协同结合导致核心 以及TRIM 5 α-Rh三聚体与病毒协同结合 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决定簇; 2）阐明 通过TRIM 5 α-rh的逆转录病毒限制中的B-box 2-B-box 2相互作用;和3）测定 TRIM 5 α-rh三聚体与HIV-1衣壳结合。完成拟议的研究将有助于我实现 我的长期目标是成为HIV-1/AIDS领域的独立调查员。为此， 丹娜-法伯癌症研究所与约瑟夫·索德罗斯基博士的实验室相结合， 环境，以实现这一目标。此外，还将举办几次培训班，以顺利完成 从授标的指导阶段过渡到独立阶段。

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.