ACTIVITY AND INHIBITION OF THE TAT PROTEIN FROM HIV

HIV TAT 蛋白的活性和抑制

• 批准号: 3143865
• 负责人: ALAN D FRANKEL
• 金额: $ 19.65万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-30 至 1994-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3143865
• 关键词: antiAIDS agent; antibody; binding proteins; cell nucleus; chelating agents; gene expression; human immunodeficiency virus; human subject; latent virus infection; metals; protein metabolism; protein purification; protein structure; receptor; synthetic peptide; tissue /cell culture; virus RNA; virus protein; virus replication

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.

人类免疫缺陷病毒(HIV)是AIDS和GT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 发展艾滋病。由于目前还没有完全有效的药物来治疗 治疗艾滋病，由于没有预防艾滋病毒感染的疫苗，艾滋病 显然，这仍然是一个主要的健康问题。 HIV编码几种在SIMPLE中找不到的调节蛋白 逆转录病毒。Tat是这些蛋白质中的一种，是HIV的强大激活剂。 基因表达，对艾滋病毒复制是必不可少的。TAT提供了一个 作为抗艾滋病药物的良好靶点，因为TAT的抑制剂 有望阻止艾滋病毒复制或防止病毒从其 潜伏期。虽然这些药物不能消除艾滋病毒的感染， 它们可以预防或减缓无症状感染的进展。 向临床艾滋病致敬。 要设计一种针对TAT的药物，重要的是要了解 更多关于TAT的结构和功能的信息。我们之前已经展示了 纯化的TAT在体外可以形成金属连接的二聚体。这 应用程序描述了旨在确定哪些活动的 TAT蛋白的形式以及金属结合如何参与TAT 功能。异常的富含半胱氨酸的金属结合可能参与了TAT 功能。不寻常的富含半胱氨酸的金属结合区可能提供 特别容易失活的目标。我们还展示了 Tat蛋白可以被组织培养中生长的细胞摄取 激活病毒启动子。确定其意义的实验 细胞摄取TAT与艾滋病的进展和逃避 HIV潜伏期被提出。 潜在的反式激活和细胞摄取的抑制剂将是 使用本申请中描述的检测方法进行测试。可能的抑制剂 包括抗TAT抗体、来自TAT可能竞争的多肽 二聚或其他TAT-蛋白质或TAT-核酸相互作用， 针对金属结合的螯合剂和其他化合物 位点，以及针对碱性区域的化合物。实验已经 旨在回答有关TAT活动的基本问题 同时测试抑制TAT的新策略。