FUNCTION AND INHIBITION OF THE REV GENE PRODUCT OF HIV

与其他逆转录病毒一样，人类病毒的整合前病毒形式 免疫缺陷病毒（HIV）产生多顺反子前体转录物 可以通过各种方式拼接来表达单个病毒基因。 的 HIV的rev基因产物是一种调控蛋白， 不同的病毒mRNA种类。 通过一个不为人知的 转录后机制，rev是表达 病毒体结构蛋白;这种突变的HIV前病毒， 功能性Rev基因不能产生感染性病毒颗粒。 旨在抑制rev功能的药物或遗传干预 可能因此提供了一种维持病毒潜伏期、阻断HIV 复制，并减缓感染者的疾病进展。 利用分子遗传学技术，我们将研究功能性 rev蛋白的结构和作用机制以及 病毒基因组中的rev反应元件（RRE），寻找潜在的 治疗干预的目标。 系统性突变的影响 将在瞬时转染试验中评价rev和RRE中的表达。 其活性可以精确控制的改进形式的rev将被 用来分析对这种蛋白质的反应的顺序。 我们 将测试源自RRE的寡核苷酸序列的能力，或 rev蛋白本身的突变形式，以阻断 使用稳定转染的报告细胞系，我们将 开发一种快速检测系统， 这一关键病毒功能的药理学抑制剂。 的信息 将为设计抗病毒基因治疗或 具体的化疗药物，也将提供深入了解 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.