VPR AND HIV INFECTION

人类免疫缺陷病毒（HIV）是导致 艾滋病是一种复杂的逆转录病毒，其控制水平由调节和 在简单的逆转录病毒中没有发现的辅助蛋白。重要的是要 我们继续研究这些调节/辅助蛋白，因为它们提供了一个 控制正常宿主细胞过程的窗口。该补助金的重点是 细胞周期进程的相互作用，导致细胞聚集在 细胞周期的G2期和随后的有丝分裂阻滞。已经 不清楚为什么这对病毒有益。 在本提案中，我们试图了解Vpr的作用机制， 解决了这赋予病毒的选择优势的问题。我们 将严格检验G2期阻滞的选择是由于以下假设： 在细胞周期的G2期增加病毒产生， 半衰期短。这将通过设置混合感染之间 会或不会引起细胞周期停滞的病毒，然后测量 在病毒的生命周期的条件下， 感染的细胞被实验性地操纵。为了了解 Vpr诱导的G2期阻滞，我们将确定Vpr和G2期阻滞之间的相互作用是否 细胞周期调节因子Cdc 25可以解释Vr对细胞周期的影响 逮捕了这将通过生化和遗传实验来确定 Vpr抑制Cdc 25活性的性质和重要性。最后我们 将表征Vpr对有丝分裂中染色体分离的影响。 预计这些结果将使我们能够更全面地了解 病毒与宿主的相互作用。

The human immunodeficiency virus (HIV), the causative agent of AIDS, is complex retrovirus with levels of control imparted by regulatory and accessory proteins not found in simple retroviruses. It is important to continue to study these regulatory/accessory proteins because they offer a window into control of normal host cell processes. This grant focuses on the interaction of the cell cycle progression by causing cells to accumulate in the G2 phase of the cell cycle and to a subsequent block in mitosis. It has been unclear why this would be beneficial for the virus. In this proposal, we seek to understand the mechanism of action of Vpr and resolve the question of the selective advantage this confers on the virus. We will critically test the hypothesis that the selection for G2 arrest is due to increased virus production in the G2 phase of the cell cycle in cells with a short half-life. This will be done by setting up mixed infections between viruses that do or do not cause cell cycle arrest, and then measuring the relative fitness of the viruses under conditions where the life-span of the infected cells is experimentally manipulated. To understand the mechanism of Vpr-induced G2 arrest, we will determine if the interaction between Vpr and the cell cycle regulators, Cdc25, can explain the effects of Vr on cell cycle arrest. This will be done with biochemical and genetic experiments to define the nature and importance of inhibition of Cdc25 activity by Vpr. Finally, we will characterize the effect of Vpr on segregation of chromosomes in mitosis. It is anticipated that these results will allow us to more fully understand the interaction of the virus with its host.