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

最近发现的猕猴蛋白；TRIM5&lt；是造成对 这些灵长类动物感染人类免疫缺陷VIMS(HIV-1)。我们已经对……的限制进行了讨论。 HIV-1通过TRIM5和rh感染，在感染过程中病毒核心在细胞质中非常稳定。这表明 通过改变进入的逆转录病毒核心的稳定性来阻断逆转录病毒的复制 宿主细胞。分开，！HIV-1衣壳实现的进入逆转录病毒核心稳定性的变化 突变也会导致受精能力的丧失；这些发现表明，病毒核心的调节 最近发现的恒河猴蛋白TRIMSalpha(TRIM5pha-rh)是导致 对这些灵长类动物感染人类免疫缺陷病毒(HIV-1)施加的限制。我们有 TRIM5α-rh对HIV-1的限制与细胞质内病毒核心稳定性降低的关系 在感染期间。这表明TRIM5α-rh蛋白通过改变逆转录病毒的 进入宿主细胞的逆转录病毒核心的稳定性。另外，进料稳定性的变化 通过HIV-1衣壳突变获得的逆转录病毒核心也会导致传染性的丧失。总而言之，这些 研究结果表明，病毒核心稳定性的调节不利于感染。值得注意的是，我们有 在TRIM5pha-rh的B-box 2区域发现了突变，该突变结合了HIV-1传入的病毒核心，但 核心的稳定性或传染性不受影响。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。出于这些原因，我们假设绑定到 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TRIM5α-rh三聚体与逆转录病毒核心的协同结合导致核心 感染的不稳定和抑制；以及TRIM5α-rh三聚体与病毒的协同结合 核心是由B-box 2-B-box 2三聚体间相互作用介导的。为了研究这一假设，我们将：1) 通过结构-功能研究确定B-box 2限制逆转录病毒的决定因素；2)阐明 B-box 2-B-box 2在TRIM5α-rh限制逆转录病毒中的相互作用；以及3)检测协同结合 TRIM5α-rh三聚体与HIV-1衣壳结合。拟议研究的完成将有助于我实现 我的长期目标是成为艾滋病毒-1/艾滋病领域的独立调查员。出于这个原因， Dana-Farber癌症研究所与Joseph Sodroski博士的实验室相结合是足够的 实现这一目标的环境。此外，还将举办几个培训班，以使 从授奖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.