HIV-1 VPR AND BASIC CELLULAR FUNCTIONS

HIV-1 VPR 和基本细胞功能

• 批准号: 2005446
• 负责人: RICHARD YUQI ZHAO
• 金额: $ 9.39万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2005446
• 关键词: Schizosaccharomyces pombe; cell cycle; enzyme activity; gene expression; gene mutation; human immunodeficiency virus 1; immunoprecipitation; molecular cloning; virus cytopathogenic effect; virus infection mechanism; virus protein; yeast two hybrid system

HIV-l Vpr blocks host cell cycle progression and interrupts cytoskeletal structures, two effects which may be related to HIV pathogenesis. At present, little is known about how Vpr affects these cellular functions and our studies are designed specifically to address this question. We have developed a novel fission yeast model system, in which we found that Vpr-induced cellular changes were very similar to those observed in human cells. As compared with human systems, fission yeast offers a simple yet genetically well-defined eukaryotic system to study mechanisms of Vpr action at both genetic and biochemical levels. We plan to use wild-type and previously characterized mutants to identify cellular pathways on which Vpr impinges and to discover the function of Vpr, something that cannot yet be done easily in mammalian cells. The fact that p34cdc2, cyclin B and other cell cycle control genes are interchangeable between human and fission yeast makes the fission yeast system relevant for studying related functions. Our goal is to use this model system l) to define domains of Vpr required to interrupt cellular functions; 2) to identify which cell cycle G2 control pathway(s) Vpr affects, and 3) to determine what gene product(s) Vpr interacts with. Our preliminary studies indicated that Vpr is likely interacting with upstream regulators of the G2-M transition but not with gene products from the DNA damage pathway. We have also identified protein candidates that bind to Vpr. In this study, we will pinpoint the cellular targets of the Vpr effects and characterize the Vpr-interacting proteins. We will further expand our studies to test the obtained results in a mammalian system. It is our expectation that the proposed studies will provide new insights into the mechanism of action of Vpr on affecting host cell functions and HIV pathogenesis.

HIV-L VPR阻断宿主细胞周期的进程并中断细胞骨架 结构，可能与HIV发病机理有关的两种作用。在 现在，关于VPR如何影响这些细胞函数的知之甚少 我们的研究专门用于解决这个问题。 我们已经开发了一种新型的裂变酵母模型系统，我们发现 VPR诱导的细胞变化与在 人类细胞。与人类系统相比，裂变酵母提供 简单但遗传上明确的真核系统研究机制 VPR作用在遗传和生化水平上的作用。我们计划使用 野生型和先前表征的突变体以鉴定细胞 VPR撞击并发现VPR功能的途径， 在哺乳动物细胞中尚不容易完成的事情。事实 P34CDC2，细胞周期蛋白B和其他细胞周期控制基因是 可在人与裂变酵母之间互换使裂变酵母 与研究相关功能相关的系统。我们的目标是使用它 模型系统l）定义中断细胞所需的VPR域 功能； 2）确定哪种细胞周期G2控制途径VPR 影响和3）确定哪种基因乘积VPR与之相互作用。我们的 初步研究表明，VPR可能正在与 G2-M过渡的上游调节剂，而不是基因产物 从DNA损伤途径。我们还确定了候选蛋白质 与VPR结合。在这项研究中，我们将确定细胞靶标 VPR效应并表征VPR相互作用蛋白。我们将 进一步扩展我们的研究以测试哺乳动物的结果 系统。我们期望拟议的研究将提供新的 对VPR影响宿主细胞的作用机理的见解 功能和HIV发病机理。