Discovery and Mechanism of Antiretroviral Factors

我们已经成功地确定了一些具有抗逆转录病毒活性的基因，并继续这样做 项目将自然而然地进展，并详细检查一个有效的行动机制 目标1中的限制因素(MX2)，以及更广泛的其他命中和其他 AIM 2中的ISG筛查。 在目标1中，我们将建立在我们的发现的基础上，通过确定MX2是一种抗逆转录病毒蛋白的机制 行动。MX2在逆转录间以衣壳依赖的方式抑制HIV-1感染 和融合。一些潜在的机制可能是这一活动的基础。首先，MX2可能直接针对 传入的病毒衣壳，以一种类似于TRIM5a和FVL抗逆转录病毒蛋白的方式，关键是，我们有 在CA中发现了许多氨基酸，当突变时，它们对MX2具有抗病毒活性。这些 将在我们的结合和核心命运分析中作为关键的特异性控制。此外，我们将确定 MX2是否与核运输成分结合，MX2核靶向信号的性质 对于其抗病毒活性以及HIV-1进入细胞核的路线是否影响其 对MX2的易感性。最终，这些实验应该确定MX2如何发挥作用以及衣壳是否 识别和/或特定的核进入途径闭塞是原因之一。 我们的ISG筛查平台已经发现了许多(-30)候选抗逆转录病毒蛋白，它们要么保护 或减少ISG表达细胞产生的感染性病毒的产量。目前，我们的分析 这些命中中的几个正在跟进的不同阶段。在目标2中，我们将遵循与我们的 以前在MX2和CNP上的工作。最终，我们将致力于准确地确定这些ISG是如何抑制 病毒复制，关键是它们是否是干扰素全面抗病毒活性所必需的。最后， 我们正在筛选最初构建的库中没有的其他ISG。再一次走上这条路 这些研究所采取的方法将反映我们以前成功的方法。 相关性(请参阅说明)： 哺乳动物，包括人类，有一系列的抗病毒防御机制。这一机制通过它 人们对这些抗病毒防御措施的了解才刚刚开始。识别和理解 抗逆转录病毒基因产物的作用机制可能导致全新的化疗 应对包括艾滋病在内的传染病的战略。

We have succeeded in identifying a number of genes with antiretroviral activity, and the continuation of this project will naturally progress with a detailed examination of the mechanism of action of one validated restriction factor (Mx2) in Aim 1, and a more broadly based follow up of additional hits, as well as additional ISG screening in Aim 2. In Aim 1 we will build on our discovery that Mx2 is an antiretroviral protein by determining its mechanism of action. Mx2 inhibits HIV-1 infection in a capsid dependent manner at a step between reverse transcription and integration. A few potential mechanisms might underlie this activity. First, Mx2 might directly target the incoming viral capsid, in a manner akin to the TRIM5a and Fvl antiretroviral proteins Crucially, we have found a number of amino acids in CA that when mutated confer resistance to Mx2 antiviral activity. These will serve as key specificity controls in our binding and core-fate assays. Additionally, we will determine whether Mx2 binds to nuclear transport components, whether the nature of the Mx2 nuclear targeting signal is important for its antiviral activity and whether the route taken by HIV-1 into the nucleus affects its susceptibility to Mx2. Ultimately these experiments should determine how Mx2 functions and whether capsid recognition and/or specific nuclear entry pathway occlusion is responsible. Our ISG screening platform has revealed numerous (-30) candidate antiretroviral proteins that either protect cells from infection, or reduce the yield of infectious virons from ISG expressing cells. Currently, our analysis of these hits is at various stages of follow up. In Aim 2 we will follow the same paths exemplified by our previous work on Mx2 and CNP. Ultimately, we will aim to determine precisely how these ISGs inhibit reroviral replication, and crucially whether they are required for the full antiviral activity of interferon. Finally, we are screening additional ISGs that were not present in our originally constructed libraries. Again the path taken by these studies will mirror our previously successful approaches. RELEVANCE (See instructions): Mammals, including humans, have an array of antiviral defense mechanisms. The mechanism by which these antiviral defenses work is only beginning to be understood. Identifying and understanding the mechanism of action of antiretroviral gene products could lead to completely new chemotherapeutic strategies for tackling infectious diseases, including AIDS.