REGULATION OF HIV GENE EXPRESSION BY TAT PEPTIDES

描述：（改编自申请人的摘要）的长期目标 本课题旨在了解达特的生化机制 （反式激活因子）作用，并利用达特功能的基本知识， 设计HIV复制的拮抗剂。 首先，要理解达特函数， 研究人员建议将重点放在一种新的人类细胞蛋白上， kDa，他已经证明（1）与达特紧密结合，（2）与 达特在体外，和（3）加强达特反式激活时，引入 啮齿动物细胞 作者将进一步表征该36 kDa蛋白， 描绘与之结合的达特识别序列， 种和细胞类型表达，克隆其cDNA并研究其在达特中的作用 反式激活 第二，在研究理解达特摄取，作者 将进一步描述他发现的一种90 kDa的细胞表面蛋白， 在体内和体外与达特结合。 他建议分析 通过交联研究，该90 kDa蛋白在达特细胞摄取中， Scatchard分析，以描绘用于结合的达特识别序列 90 kDa，并通过氨基酸序列分析确定90 kDa是否 kDa是一种新的蛋白质。 如果有必要，他将分离出一个cDNA克隆， 90 kDa，用于进一步研究。 三是在研究上进一步发展达特 HIV复制的肽拮抗剂，研究者将利用 （1）细胞的达特识别序列的映射研究结果 摄取，（2）TAR结合和（3）36 kDa结合，试图设计 小的达特肽将被细胞吸收并阻止艾滋病毒 复制的 作为一个长期目标，有前途的达特肽的类似物 将通过肽模拟技术合成， 有效的达特拮抗剂。 第四，探索利用达特肽 将外源多肽转运到细胞中的序列， 建议分析细胞摄取，核运输，和生物 含有达特“递送”序列的融合多肽的功能。 他 建议首先检查116个氨基酸的HIV Rev（病毒体调节因子 蛋白质表达[以前为art/trs]）与达特融合的蛋白质。

DESCRIPTION: (Adapted from applicant's abstract) The long-term goals of this project are to understand the biochemical mechanism of Tat (transactivator) action and to utilize basic knowledge of Tat function to design antagonists of HIV replication. First, to understand Tat function, the investigator proposes to focus on a new human cellular protein of 36 kDa that he has shown to (1) bind tightly to Tat, (2) form a complex with Tat in vitro, and (3) potentiate Tat transactivation when introduced into rodent cells. The author will further characterize this 36 kDa protein, delineate the Tat recognition sequences for binding it, determine cell species and cell type expression, clone its cDNA and study its role in Tat transactivation. Second, in studies to understand Tat uptake, the author will further characterize a 90 kDa cell surface protein that he has found to bind to Tat in vivo and in vitro. He proposes to analyze the role of this 90 kDa protein in Tat cellular uptake by cross-linking studies and Scatchard analysis, to delineate the Tat recognition sequences for binding 90 kDa, and to determine by amino acid sequence analysis whether the 90 kDa is a new protein. If warranted, he will isolate a cDNA clone encoding 90 kDa for further studies. Third, in studies to develop further Tat peptide antagonists of HIV replication, the investigator will utilize the results of mapping studies of Tat recognition sequences for (1) cell uptake, (2) TAR binding, and (3) 36 kDa binding in attempts to design small Tat peptides that will be taken up by cells and block HIV replication. As a long-range goal, analogues of promising Tat peptides will be synthesized by peptide mimetic technology to yield stable and more efficient Tat antagonists. Fourth, to explore the use of Tat peptide sequences to transport foreign polypeptides into cells, the author proposes to analyze the cellular uptake, nuclear transport, and biological function of fusion polypeptides containing Tat "delivery" sequences. He proposes to first examine the 116 amino acid HIV Rev (regulator of virion protein expression [formerly art/trs]) protein fused to Tat.