ACTIVITY AND INHIBITION OF THE TAT PROTEIN FROM HIV

人类免疫缺陷病毒（HIV）是艾滋病的病原体。 据估计，目前至少有200万人 仅在美国就感染了艾滋病毒，预计大多数人将 发展成艾滋病。 由于目前还没有完全有效的药物， 治疗艾滋病，由于没有疫苗可以预防艾滋病毒感染，艾滋病 显然仍是一个重大的健康问题。 HIV编码了几种调节蛋白， 逆转录病毒 其中一种蛋白质达特是艾滋病病毒的一种强有力的激活剂 基因表达，是艾滋病毒复制所必需的。 达特提供了一个 这是抗艾滋病药物的一个很好的靶点，因为达特的抑制剂 预期可阻断HIV复制或防止诱导病毒从其 潜伏状态 虽然这些药物不能消除艾滋病毒感染， 它们可以预防或减缓无症状感染的进展， 临床艾滋病 为了设计出一种针对达特的药物， 更多关于达特的结构和功能。 我们先前已经表明 纯化的达特可以在体外形成金属连接的二聚体。 这 应用程序描述了设计用于确定活性物质 达特蛋白的形式以及金属结合如何参与达特 功能 这种不寻常的富含半胱氨酸的金属结合可能与达特有关 功能 不寻常的富含半胱氨酸的金属结合区可能提供了一个新的机制。 特别容易失活的目标。 我们还表明 达特蛋白可以被组织培养中生长的细胞吸收， 激活病毒启动子 确定显著性的实验 达特的细胞摄取的进展，艾滋病和逃避 艾滋病毒潜伏期的建议。 反式激活和细胞摄取的潜在抑制剂将是 使用本申请中所述的测定进行测试。 可能的抑制剂 包括抗Tat抗体、来自达特的肽，其可能竞争 二聚化或其它TAT-蛋白质或TAT-核酸相互作用， 螯合剂和其它针对金属结合的化合物 位点和针对碱性区域的化合物。 实验 旨在回答有关达特活动的基本问题， 同时测试抑制达特的新策略。

The human immunodeficiency virus (HIV) is the etiologic agent of AIDS> It is estimated that at least 2 million individuals are presently infected with HIV in the United States alone and most are expected to develop AIDS. Since there is currently no completely effective drug for treating AIDS, and since no vaccine exists to prevent HIV infection, AIDS clearly remains a major health problem. HIV encodes several regulatory proteins that are not found in simpler retroviruses. One of these proteins, tat is a powerful activator of HIV gene expression and is essential for HIV replication. Tat provides a good target for an anti-AIDS drug since inhibitors of tat would be expected to block HIV replication or prevent induction of virus from its latent state. Although such drugs would not eliminate infection by HIV, they may prevent or slow down the progression from asymptomatic infection to clinical AIDS. To design a drug directed against tat, it will be important to understand more about the structure and function of tat. We have previously shown that purified tat can form metal-linked dimers in vitro. This application describes experiments designed to determine what the active form of the tat protein is and how metal-binding may be involved in tat function. The unusual cysteine-rich metal-binding may be involved in tat function. The unusual cysteine-rich metal-binding region may provide a particularly susceptible target for inactivation. We have also shown that the tat protein can be taken up by cells grown in tissue culture and activate the viral promoter. Experiments to determine the significance of cellular uptake of tat for the progression of AIDS and for escape from HIV latency are proposed. Potential inhibitors of transactivation and cellular uptake will be tested using assays described in this application. Possible inhibitors include anti-tat antibodies, peptides from tat which might compete for dimerization or other tat-protein or tat-nucleic acid interactions, chelating agents and other compounds directed against the metal-binding sites, and compounds directed against the basic region. Experiments have been designed to answer basic questions about tat activity while at the same time testing new strategies for tat inhibition.