FUNCTION AND INHIBITION OF THE REV GENE PRODUCT OF HIV

像其他逆转录病毒一样，人类的整合前病毒形式 免疫缺陷病毒(HIV)产生一个多顺反子前体转录本 它们可以以不同的方式拼接起来，以表达单个病毒基因。这个 HIV的REV基因产物是一种调控蛋白的表达 病毒基因的不同种类。通过一个鲜为人知的 转录后机制，REV是需要的表达 病毒粒子结构蛋白；这种突变的艾滋病毒前病毒缺乏一种 功能性rev基因不能产生具有感染性的病毒颗粒。 旨在抑制REV功能的药物或遗传干预 可能因此提供了一种保持病毒潜伏期、阻断HIV的方法 复制，并减缓感染者的疾病进展。 利用分子遗传学技术，我们将研究功能 REV蛋白和A蛋白的结构和作用机制 病毒基因组中的REV反应元件(RRE)，寻找潜在的 治疗干预的靶点。系统性突变的影响 在REV和RRE中，将在瞬时转染试验中进行评估。 其活动可以精确控制的修改形式的rev将是 用来分析对这种蛋白质的反应中的一系列事件。我们 将测试从RRE衍生的寡核苷酸序列的能力，或 突变形式的rev蛋白本身，以阻止 野生型Rev.使用稳定转染的报告细胞系，我们将 开发一种可用于筛选潜力的快速检测系统 这一关键病毒功能的药物抑制剂。这些信息 将在设计抗病毒基因疗法或 特定的化疗药物，并将提供对 神秘的转录后机制，控制细胞以及 病毒，基因表达。

Like other retroviruses, the integrated proviral form of the human immunodeficiency virus (HIV) produces a multicistronic precursor transcript that can be spliced in various ways to express individual viral genes. The rev gene product of HIV is a regulatory protein that controls expression of the different viral mRNA species. Acting through a poorly-understood post-transcriptional mechanism, rev is required for the expression of virion structural proteins; this, mutant HIV provirus that lack a functional rev gene fail to produce infectious viral particles. Pharmacologic or genetic interventions aimed at inhibiting rev function might therefore provide a means to maintain viral latency, block HIV replication, and slow the progression of disease in infected persons. Using techniques of molecular genetics, we will investigate the functional architecture and mechanism of action of the rev protein and of a rev-responsive element (RRE) in the viral genome, searching for potential targets for therapeutic intervention. The effects of systematic mutations in rev and in the RRE will be evaluated in a transient-transfection assay. Modified forms of rev whose activity can be precisely controlled will be used to dissect the sequence of events in the response to this protein. We will test the ability of oligonucleotide sequences derived from the RRE, or of mutants forms of the rev protein itself, to block the activity of wild-type rev. Using stably-transfected reporter cell lines, we will develop a rapid assay system that could be used to screen potential pharmacologic inhibitors of this critical viral function. The information to be gained will be of value in designing antiviral gene therapy or specific chemotherapeutic agents, and will also provide insight into the enigmatic post-transcriptional mechanisms that control cellular, as well as viral, gene expression.