VPR AND HIV INFECTION

描述(改编自研究人员摘要)：获得性免疫 缺乏综合症(艾滋病)的特征是进行性消耗。 CD_4淋巴细胞。艾滋病的病原体，人类 免疫缺陷病毒(HIV)是一种细胞性逆转录病毒。尽管 HIV感染者体内CD4细胞丢失的机制尚不清楚， 最近的数据表明，艾滋病毒对感染细胞的直接杀伤作用 这很可能是它自己造成的。HIV的一个辅助基因，VPR基因， 在艾滋病毒的细胞病变效应中起着重要作用。在 存在完整的vpr基因，感染细胞的培养几乎 完全死亡，而感染了一种病毒的培养物 突变的vpr基因存活和复制的动力学与 未受感染的细胞。那些在感染了野生型病毒后存活的细胞 病毒含有带有突变的vpr基因。此外，vpr的表达 它本身通过使细胞积累在 VPR在病毒生命周期中也扮演着另一个角色，它允许 进入感染细胞细胞核的整合前复合体 在有丝分裂之前。这一特性使艾滋病毒能够最终感染 分化的巨噬细胞，体内病毒的重要储存库。 在本申请中，我们试图了解的作用机制 VPR。最终，我们希望确定VPR是否在 CD4细胞在艾滋病进展过程中的下降。具体地说，我们将使用 VPR突变和定点突变的基因筛查 确定VPR不同功能之间的相关性。 这将使我们能够对vpr在艾滋病毒中的作用进行建模。 发病机制。我们还将描述VPR和VPX的影响 HIV-2对细胞增殖的影响。接下来，我们将描述细胞周期的特征 阶段(S)受VPR影响，并分析正常关系 细胞周期调控和VPR功能。然后我们将确定 VPR在原代细胞中的作用，并具体确定VPR如何 影响淋巴细胞和巨噬细胞的半衰期。最后，我们 将使用筛子来抑制酵母中VPR的毒性作用 确定VPR的宿主细胞靶点。在此应用程序中，我们寻求 了解VPR的作用机制。最终，我们希望 确定VPR是否在艾滋病期间CD4细胞的下降中起作用 进步。具体地说，我们将使用VPR的基因筛查 突变以及定点突变来确定 VPR不同功能之间的相关性。这将使我们能够 建立vpr在HIV致病机制中的作用模型。我们还将 表征HIV-2的VPR和VPX对细胞增殖的影响。 接下来，我们将表征VPR对细胞周期阶段(S)的影响，以及 分析正常细胞周期调控与vpr的关系 功能。然后我们将确定VPR在原代细胞中的影响， 并具体确定VPR如何影响半衰期 淋巴细胞和巨噬细胞。最后，我们将使用一个屏幕来 酵母中VPR毒性作用的抑制物鉴定宿主细胞 VPR的目标。

DESCRIPTION (Adapted from investigator's abstract): The acquired immune deficiency syndrome (AIDS) is characterized by a progressive depletion of CD4 lymphocytes. The causative agent of AIDS, the human immunodeficiency virus (HIV) is a cytopathic retrovirus. Although the mechanisms of CD4 cell loss in HIV-infected people remains unclear, recent data indicate the direct cell killing of infected cells by HIV itself is likely responsible. An accessory gene of HIV, the vpr gene, plays an important role in the cytopathic effect of HIV. In the presence of an intact vpr gene, cultures of infected cells are nearly completely killed, while cultures infected with a virus that has a mutated vpr gene survive and replicate with the same kinetics as uninfected cells. Those cells that survive infection with a wild-type virus contain vpr genes with mutations. Furthermore, expression of Vpr itself alters cell cycle progression by causing cells to accumulate in G2/M. Vpr also plays another role in the virus life-cycle by allowing the pre-integration complex to enter the nucleus of infected cells before mitosis. This property allows HIV to infect terminally differentiated macrophages, an important reservoir of virus in the body. In this application, we seek to understand the mechanism of action of Vpr. Ultimately, we hope to determine if Vpr plays a role in the decline of CD4 cells during AIDS progression. Specifically, we will use a genetic screen for vpr mutations as well as site-directed mutations to determine the correlation between the different functions of Vpr. This will allow us to make models of the roles of Vpr in HIV pathogenesis. We will also characterize the effects of Vpr and Vpx of HIV-2 on cell proliferation. Next, we will characterize the cell cycle stage(s) affected by Vpr, and analyze the relationship between normal cell cycle controls and Vpr function. We will then determine the effects of Vpr in primary cells, and specifically determine how Vpr influences the half-life of lymphocytes and macrophages. Finally, we will use a screen for suppressors of the toxic effects of Vpr in yeast to Identify host cell targets of Vpr. In this application, we seek to understand the mechanism of action of Vpr. Ultimately, we hope to determine if Vpr plays a role in the decline of CD4 cells during AIDS progression. Specifically, we will use a genetic screen for vpr mutations as well as site-directed mutations to determine the correlation between the different functions of Vpr. This will allow us to make models of the roles of Vpr in HIV pathogenesis. We will also characterize the effects of Vpr and Vpx of HIV-2 on cell proliferation. Next, we will characterize the cell cycle stage(s) affected by Vpr, and analyze the relationship between normal cell cycle controls and Vpr function. We will then determine the effects of Vpr in primary cells, and specifically determine how Vpr influences the half-life of lymphocytes and macrophages. Finally, we will use a screen for suppressors of the toxic effects of Vpr in yeast to Identify host cell targets of Vpr.