VPR AND HIV INFECTION

描述（改编自研究者摘要）：获得性免疫 缺乏综合征（艾滋病）的特点是进行性消耗 CD 4淋巴细胞艾滋病的病原体，人类 免疫缺陷病毒（HIV）是一种致细胞病变的逆转录病毒。虽然 HIV感染者中CD 4细胞损失的机制仍不清楚， 最近的数据表明，HIV直接杀死受感染的细胞， 可能是自己的责任。艾滋病病毒的辅助基因vpr基因， 在HIV的细胞病变效应中起重要作用。在 当存在完整的vpr基因时，感染细胞的培养物几乎 完全杀死，而文化感染的病毒， 突变的vpr基因以与 未感染的细胞那些在感染野生型病毒后存活的细胞 病毒含有突变的vpr基因。此外，Vpr的表达 它本身通过使细胞在细胞周期中积累来改变细胞周期进程， G2/M。Vpr在病毒的生命周期中还扮演着另一个角色， 整合前复合物进入受感染细胞的细胞核 在有丝分裂之前。这种特性使艾滋病病毒能够感染晚期 分化的巨噬细胞，体内重要的病毒储存库。 在本申请中，我们寻求理解以下的作用机制： Vpr。最终，我们希望确定Vpr是否在 在艾滋病发展过程中，CD 4细胞减少。具体来说，我们将使用 对vpr突变和定点突变进行基因筛查 以确定Vpr的不同函数之间的相关性。 这将使我们能够制作Vpr在艾滋病毒中作用的模型 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 受Vpr影响的分期，并分析正常 细胞周期控制和Vpr功能。然后我们将确定 Vpr在原代细胞中的作用，并特别确定Vpr如何 影响淋巴细胞和巨噬细胞的半衰期。最后我们 将在酵母中筛选Vpr毒性作用的抑制剂 以鉴定Vpr的宿主细胞靶点。在本申请中，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CD 4细胞的下降中起作用 进展具体地说，我们将使用基因筛查vpr 突变以及定点突变，以确定 Vpr的不同功能之间的相关性。这将使我们 建立Vpr在HIV发病机制中作用的模型。我们还将 表征HIV-2的Vpr和Vpx对细胞增殖的影响。 接下来，我们将表征受Vpr影响的细胞周期阶段， 分析正常细胞周期控制与Vpr之间的关系 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。最后，我们将使用屏幕 Vpr对酵母细胞毒性作用的抑制因子 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.