Understanding Retroviral Gag and RNA Targeting and Virus Assembly

我们研究了产生传染性病毒所需的复杂相互作用。我们正在研究艾滋病毒-1大分子的贩运和组装。一旦从细胞核输出，HIV-1RNA需要运输到适当的亚细胞位置进行翻译或包装；同样，新合成的Gag需要靶向质膜进行病毒组装。使用我们新开发的成像工具，我们已经确定HIV-1RNA使用扩散作为主要的运输机制。HIV-1 RNA运输到质膜进行组装；我们将调查这种靶向事件的性质，包括在组装过程中是以单体RNA还是二聚体RNA为靶点，以及RNA在质膜附近的停留时间。我们目前正在验证这样的假设，即Gag-二聚体RNA相互作用是逆转录病毒组装的成核点。我们正在研究极化T细胞和病毒突触中的GAG和RNA靶向。这些实验将提供对HIV-1复制方面的了解，而我们目前对这些方面知之甚少，并将使我们更好地了解HIV-1是如何利用细胞机械来运输其大分子的。[对应于2011年10月艾滋病毒耐药计划现场访问报告中的HU项目3,2015年更名为艾滋病毒动态和复制计划]

We have examined the complex interactions needed for generating an infectious virus. We are studying the trafficking of HIV-1 macromolecules and assembly. Once exported from the nucleus, HIV-1 RNA needs to traffic to proper subcellular locations to be translated or packaged; similarly, the newly synthesized Gag needs to be targeted to the plasma membrane for virus assembly. Using our newly developed imaging tools, we have determined that HIV-1 RNA uses diffusion as the major mechanism for transport. HIV-1 RNA traffics to the plasma membrane for assembly; we will investigate the properties of this targeting event, including whether monomeric or dimeric RNA is targeted during assembly and the dwell time of the RNA that stays near the plasma membrane. We are currently testing the hypothesis that Gag-dimeric RNA interaction is the nucleation point of retroviral assembly. We are investigating Gag and RNA targeting in polarized T cells and in viral synapses. These experiments will provide an understanding of aspects of HIV-1 replication that we currently know very little about and will allow us to better understand how HIV-1 uses the cellular machinery to traffic its macromolecules. [Corresponds to Hu Project 3 in the October 2011 site visit report of the HIV Drug Resistance Program, renamed the HIV Dynamics and Replication Program in 2015]