CELLULAR CONTROL OF HIV EXPRESSION

HIV表达的细胞控制

• 批准号: 3147036
• 负责人: B ROBERT FRANZA
• 金额: $ 15.7万
• 依托单位: COLD SPRING HARBOR LABORATORY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1994-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3147036
• 关键词: DNA binding protein; Escherichia coli; antisense nucleic acid; genetic transcription; human immunodeficiency virus 1; laboratory mouse; laboratory rabbit; molecular cloning; polymerase chain reaction; protein purification; protein structure function; provirus; tissue /cell culture; virus replication

This proposal addresses the definition of cellular mechanisms involved in the transcriptional activation of the HIV-1 provirus. Our previous studies have resulted in the identification of a complex set of cellular proteins that interact with control elements within the HIV-1 long terminal repeat (LTR). We have identified inducible and cell type specific proteins that interact with the duplicated kappaB site in the LTR. We have also established a microscale in vitro transcription assay to study the biochemical properties of extracts from cells at different growth stages to assess the transcriptional activity of the HIV-1 LTR and how different control elements within the LTR contribute to that activity. We are now in a position to use what we have learned and the systems that we have operational to determine if certain of the proteins interacting with the kappaB sites in the HIV-1 LTR are necessary for transcriptional activation of resident, integrated provirus. We will focus on a thorough characterization of one of the kappaB binding proteins, Rel (HIVEN86A) because we have demonstrated it to be inducible and because both the CDNA and various immunologic reagents are now available to permit structure and function analysis of the protein. We will use the anti-sense oligonucleotide approach to disrupt the production of Rel and determine the effects in cells that have the HIV-1 provirus stably integrated. We will also study the effects of over expression of Rel or disruption of expression of Rel in cells acutely infected with HIV-1. We will continue to characterize and to develop reagents for the study of the other kappaB binding proteins especially the proteins we have demonstrated to be cell type specific.

这项建议涉及细胞机制的定义 HIV-1前病毒的转录激活。 我们以前的 研究已经鉴定出一组复杂的细胞 与HIV-1长链内的控制元件相互作用的蛋白质 末端重复序列（LTR）。 我们已经确定了诱导型和细胞类型 特异性蛋白质与重复的kappaB位点相互作用， LTR 我们还建立了一个微型的体外转录实验 研究不同温度下细胞提取物的生化特性， 评估HIV-1 LTR转录活性的生长阶段 以及LTR中的不同控制元素如何对此做出贡献 活动 我们现在能够利用我们所学到的东西， 我们的系统可以用来确定某些蛋白质 与HIV-1 LTR中的kappaB位点相互作用是必要的， 转录激活的居民，整合前病毒。 我们将集中在一个彻底的特点之一卡帕B 结合蛋白，Rel（HIVEN 86 A），因为我们已经证明它是 诱导的，并且因为cDNA和各种免疫试剂都是 现在可用于允许蛋白质的结构和功能分析。 我们将使用反义寡核苷酸方法来破坏 产生Rel，并确定在具有HIV-1的细胞中的作用。 前病毒稳定整合。 我们还将研究过 细胞中Rel表达或Rel表达的急性破坏 感染了HIV-1。 我们将继续表征和开发用于研究的试剂 其他kappaB结合蛋白，特别是我们现有的 证明是细胞类型特异性的。