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CELLULAR CONTROL OF HIV EXPRESSION

HIV表达的细胞控制

基本信息

批准号：
3147039
负责人：
B FRANZA
金额：
$ 17.12万
依托单位：
COLD SPRING HARBOR LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-09-30 至 1994-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3147039
关键词：
DNA binding protein Escherichia coli antisense nucleic acid genetic transcription host organism interaction human immunodeficiency virus 1 laboratory mouse laboratory rabbit molecular cloning polymerase chain reaction protein purification protein structure function provirus tissue /cell culture virus replication

项目摘要

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.

该提案涉及对所涉及的细胞机制的定义在HIV-1前病毒的转录激活中。我们以前的研究发现了一组复杂的细胞与HIV-1Long中的控制元件相互作用的蛋白质终端重复(LTR)。我们已经确定了诱导型和细胞型与复制的kappaB位点相互作用的特定蛋白质 Ltr.我们还建立了一种微尺度的体外转录实验研究不同温度下细胞提取液的生化性质用于评估HIV-1 LTRs转录活性的生长阶段以及LTR中的不同控制元素对此有何影响活动。我们现在能够利用我们所学到的和我们可以运行的系统来确定某些蛋白质是否与HIV-1 LTR中的kappaB位点相互作用对于驻留的整合前病毒的转录激活。我们将专注于对其中一种kappab的全面描述结合蛋白，Rel(HIVEN86A)，因为我们已经证明它是诱导的，因为CDNA和各种免疫试剂都是现在可用于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干扰产生REL并确定对携带HIV-1的细胞的影响前病毒稳定整合。我们还将研究 Rel在细胞中的急性表达或表达中断感染了HIV-1病毒。我们将继续为这项研究确定特征并开发试剂其他kappaB结合蛋白，特别是我们拥有的蛋白被证明是特定细胞类型的。