Understanding Retroviral Gag and RNA Targeting and Virus Assembly

我们已经研究了产生传染性病毒所需的复杂相互作用。我们发现，来自两种不同慢病毒HIV-1和SIVmac的成熟衣壳蛋白可以组装成感染性颗粒的成熟核心。此外，我们量化了HIV-1感染过程中所需的功能性核衣壳（NC）和p6蛋白的量，并确定了用次优浓度的蛋白酶抑制剂治疗HIV-1对病毒蛋白加工和RNA成熟的影响。我们将研究HIV-1大分子的贩运和组装。一旦从细胞核输出，HIV-1 RNA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，我们将研究HIV-1 RNA在细胞质中转运的机制。HIV-1 RNA运输到质膜进行组装;我们将研究这种靶向事件的性质，包括组装过程中是否靶向单体或二聚体RNA以及RNA在质膜附近的停留时间。我们还将研究极化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复制方面的理解，我们目前对此知之甚少，并将使我们更好地了解HIV-1如何使用细胞机制来运输其大分子。[对应于2011年10月艾滋病毒耐药性项目现场访问报告中的Hu项目3]

We have examined the complex interactions needed for generating an infectious virus. We showed that the mature capsid proteins from two different lentiviruses, HIV-1 and SIVmac, can assemble into the mature core of the infectious particle. Additionally, we quantified the amounts of functional nucleocapsid (NC) and p6 protein needed during HIV-1 infection, and determined the effects of treating HIV-1 with suboptimal concentrations of protease inhibitors on viral protein processing and RNA maturation. We will study 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 will investigate the mechanisms used by HIV-1 RNA for transport in the cytoplasm. 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 will also investigate 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]