Regulatuion of HIV-1 Gene Expression in Latency by YY1, RuvB2, and ZAP

描述（由申请方提供）：使用高效抗逆转录病毒治疗（HAART），可以控制HIV-1复制，以维持血浆病毒RNA水平低于常规方法检测水平。然而，沉默的HIV-1前病毒DNA在记忆性CD 4 + T细胞中的持续存在是艾滋病患者根除病毒的主要障碍。据报道，HIV-1前病毒在这些细胞中保持沉默状态的机制有多种。据信，对活跃病毒复制的主要阻断是在转录水平。然而，在长期HAART治疗的患者中可以通过灵敏的检测方法检测到病毒RNA，这表明转录控制可能不是维持病毒潜伏期的唯一机制。该项目现在旨在了解由三个调节系统介导的HIV-1基因表达的转录后控制：无义介导的衰变（NMD）系统;我们最近鉴定的HIV翻译阻遏物（RVB 2）;和锌指抗病毒蛋白（ZAP），其沉默并触发许多病毒RNA的降解。我们将专门探讨这些系统中的每一个在维持HIV-1潜伏期中的可能作用。我们将检查HIV-1 RNA和蛋白质表达水平的CD 4阳性记忆T细胞从HIV-1感染的患者后，操纵这些调控系统的功能，使用siRNA敲低技术。发现控制HIV-1基因表达的转录后调节的主要机制是释放抑制和激活病毒复制的重要第一步。这种激活将允许通过免疫系统检测病毒感染的细胞，并通过靶向在记忆T细胞中和记忆T细胞表面上表达的病毒蛋白的疗法促进感染细胞的清除。

DESCRIPTION (provided by applicant): With Highly Active AntiRetroviral Therapy (HAART), HIV-1 replication can be controlled to maintain a plasma level of viral RNA below detection by conventional means. However, the continued presence of silent HIV-1 proviral DNA in memory CD4+ T cells is a major barrier to the eradication of the virus in AIDS patients. Multiple mechanisms have been reported by which HIV-1 proviruses are maintained in a silent state in these cells. The major block to active viral replication is believed to be at the transcriptional level. However, viral RNAs can be detected in long-term HAART-treated patients by sensitive detection methods, suggesting that transcriptional control may not be the only mechanism underlying the maintenance of viral latency. This project now aims to understand the post-transcriptional control of HIV-1 gene expression mediated by three regulatory systems: the Nonsense Mediated Decay (NMD) system; a repressor of HIV translation (RVB2) which we have recently identified; and the Zinc- finger Antiviral Protein (ZAP) which silences and triggers degradation of many viral RNAs. We will specifically explore the possible role of each of these systems in the maintenance of HIV-1 latency. We will examine the levels of HIV-1 RNA and protein expression in CD4-positive memory T cells from HIV-1 infected patients after manipulating the functions of each of these regulatory systems using siRNA knockdown technology. Discovery of the major mechanisms controlling post-transcriptional regulation of HIV-1 gene expression is an essential first step toward the release of repression and the activation of viral replication. This activation will permit the detection of virus-infected cells y the immune system and facilitate the clearance of infected cells by therapies that target viral proteins expressed in and on the surface of memory T cells.