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，对 病毒的复制。定位于受感染细胞的细胞核内， REV通过一种鲜为人知的机制来允许细胞质 某些未完全剪接的HIV-1 mRNAs的积累将 否则将被隔离在原子核中。因为这些mRNA编码 对于主要的病毒结构蛋白，HIV-1前病毒缺乏一个 功能性REV基因不能产生新的感染性病毒粒子。Rev是 因此，人们之所以感兴趣，不仅是因为它前所未有的监管 它不仅有效，而且也是抗逆转录病毒治疗的一个有希望的靶点。 利用分子遗传学技术，我们正在研究 REV的功能结构和作用机制，寻找 潜在的抑制其活性的手段。突变被引入 系统研究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用 瞬时转染法。我们的研究已经确定，现在也将 进一步鉴定，三个关键的多肽结构域 REV单体之间的相互作用，与可能的细胞 辅因子，并与靶RNA结合。每个地区的基本特征 将详细绘制成图，为合理设计提供依据 抑制剂，我们将在类REV中搜索等效域 来自其他逆转录病毒的蛋白质和选定的细胞蛋白质。我们 也将继续观察到这些区域的修饰形式可以 阻断野生型REV的活性，作为一种可能的基因治疗方法。 使用改变了靶标特异性的REV融合蛋白，我们将 剖析病毒这一途径的最低要求 活体内激活。我们还将调查 受感染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无价的 设计新的药物或遗传策略来抑制REV， 它可以用来维持HIV-1的潜伏期，减缓 在受感染的个体中的疾病。

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.