FUNCTION AND INHIBITION OF THE REV GENE PRODUCT OF HIV

人类免疫缺陷病毒1型（HIV-1）的Rev反式激活因子是 一种转录后调节蛋白， 病毒的复制。 定位于受感染细胞的细胞核内， Rev通过一种知之甚少的机制发挥作用， 某些不完全剪接的HIV-1 mRNA的积累， 否则就会被隔离在细胞核中。 因为这些基因编码 对于主要的病毒结构蛋白，HIV- 1前病毒缺乏 功能性Rev基因不能产生新的感染性病毒体。 Rev是 不仅因为其前所未有的监管， 效果，而且也是抗逆转录病毒治疗的一个有前途的目标。 利用分子遗传学技术，我们正在研究 功能结构和作用机制的Rev，寻找 抑制其活性的潜在方法。 引入突变 系统地转化为Rev及其在病毒中的顺式作用靶元件 基因组，这些突变的影响，然后使用 瞬时转染测定。 我们的研究已经确定，现在将 进一步表征，三个关键的肽结构域， Rev单体彼此之间的相互作用，与假定的细胞 辅因子和靶RNA。 每个区域的基本特征 将详细绘制，为合理设计 抑制剂，我们将在Rev样 其他逆转录病毒的蛋白质和选定的细胞蛋白质。 我们 我也将继续观察，这些区域的修改形式可以 阻断野生型Rev的活性，作为一种可能的基因治疗方法。 使用具有改变的靶特异性的Rev融合蛋白，我们将 剖析这种病毒传播途径的最低要求， 体内反式激活。 我们还将研究 受感染的T淋巴细胞的生理状态可以调节Rev活性。 从这些研究中获得的信息将是非常宝贵的， 设计抑制Rev的新的药理学或遗传学策略， 它可以用来维持HIV-1的潜伏期，减缓HIV-1的进展。 感染者的疾病。

The Rev transactivator of human immunodeficiency virus type 1 (HIV-1) is a posttranscriptional regulatory protein that is essential for replication of the virus. Localized within the nuclei of infected cells, Rev acts through a poorly understood mechanism to allow cytoplasmic accumulation of certain incompletely spliced HIV-1 mRNAs that would otherwise remain sequestered in the nucleus. Because these mRNAs code for the major viral structural proteins, HIV- 1 proviruses that lack a functional rev gene are unable to produce new infectious virions. Rev is therefore of interest not only because of its unprecedented regulatory effect, but also as a promising target for antiretroviral therapy. Using techniques of molecular genetics, we are investigating the functional architecture and mechanism of action of Rev, searching for potential means of inhibiting its activity. Mutations are introduced systematically into Rev and its cis-acting target element in the viral genome, and the effects of these mutations are then characterized using a transient transfection assay. Our studies have identified, and will now further characterize, three critical peptide domains that mediate interactions of Rev monomers with each other, with putative cellular cofactors, and with the target RNAs. Essential features of each region will be mapped in detail to provide a basis for rational design of inhibitors, and we will search for equivalent domains in Rev-like proteins from other retroviruses and in selected cellular proteins. We will also pursue the observation that modified forms of these regions can block activity of wild-type Rev, as a possible approach to gene therapy. Using Rev fusion proteins that have altered target specificity, we will dissect the minimal requirements for this pathway of viral transactivation in vivo. We will also investigate how changes in the physiologic state of an infected T lymphocyte can modulate Rev activity. The information to be gained in these studies will be invaluable in designing novel pharmacologic or genetic strategies for inhibiting Rev, which could be used to maintain HIV-1 latency and slow the progression of disease in infected individuals.