Molecular gene expression reprogramming by HIV TAT

HIV TAT 的分子基因表达重编程

• 批准号: 6893202
• 负责人: ANNA ALDOVINI
• 金额: $ 41.36万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-01 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6893202
• 关键词: Adenoviridae; DNA binding protein; HIV infections; RNA binding protein; Retroviridae; chromatin immunoprecipitation; dendritic cells; gel mobility shift assay; gene expression; gene induction /repression; gene interaction; genetic mapping; genetic recombination; green fluorescent proteins; host organism interaction; human tissue; microarray technology; molecular biology information system; polymerase chain reaction; regulatory gene; tissue /cell culture; transcription factor; transfection /expression vector; translation factor; virus infection mechanism; virus replication

DESCRIPTION (provided by applicant): The goal of this proposal is to gain insights into how HIV-1 Tat mediates its effects on host cell gene expression. Tat is among the first genes expressed during HIV-1 infection and is essential for viral gene expression and virus production. Tat increases HIV-1 gene expression by functioning as an elongation factor and interacting with TAR, a RNA sequence present at the beginning of the HIV viral transcripts, and with the host cell factors CDK9 and cyclin T1. The interaction of Tat with these and other host cell transcriptional regulators might be expected to affect host cell gene expression. Indeed, there is considerable evidence that Tat can affect the physiology of T lymphocytes, neurons and antigen presenting cells. Recent studies from our laboratory have shown that the gene expression programs of dendritic cells and macrophages are modified by pathogen exposure and that these modifications can provide important clues to pathogenesis. We have shown that dendritic cell reprogramming by HIV-1 can create conditions that favor virus spread, and that the effect is mediated by the HIV-1 transactivator Tat. This proposal is designed to improve our understanding of the means by which Tat expression causes changes in host cell gene expression in immune cells targeted by HIV-1. To accomplish this, the specific aims of the proposal are 1) To develop a library of adenovirus and retroviral vectors expressing multiple wild type and mutated Tat proteins. 2) To determine the effects of two wild type alleles of Tat on host cell gene expression in immune cells targeted by HIV-1 infection. 3) To map the domain of Tat that is responsible for modifying the host cell gene expression program. 4) To investigate the genome-scale location of the transcription factors STAT1 and IRF7 during Tat expression, as these factors are upregulated by HIV-1 infection and Tat expression. 5) To investigate the mechanism by which Tat reprograms host cell gene expression using genome-scale location analysis experiments designed to detect DNA and RNA binding.

描述（由申请人提供）：该提案的目的是了解HIV-1 TAT如何介导其对宿主细胞基因表达的影响。 TAT是HIV-1感染期间最早表达的基因之一，对于病毒基因表达和病毒的产生至关重要。 TAT通过充当伸长因子并与TAR相互作用（在HIV病毒转录物的开头以及宿主细胞因子CDK9和Cyclin T1）相互作用来增加HIV-1基因表达。 TAT与这些宿主细胞转录调节剂的相互作用可能会影响宿主细胞基因的表达。确实，有大量证据表明，TAT可以影响T淋巴细胞，神经元和抗原呈递细胞的生理。我们实验室的最新研究表明，树突状细胞和巨噬细胞的基因表达程序是通过病原体暴露来改变的，并且这些修饰可以为发病机理提供重要的线索。我们已经表明，通过HIV-1重编程的树突状细胞可以产生有利于病毒扩散的疾病，并且该作用是由HIV-1反式激活剂TAT介导的。该建议旨在提高我们对TAT表达在HIV-1靶向的免疫细胞中引起宿主细胞基因表达变化的平均值的理解。为此，该提案的具体目的是1）开发表达多种野生型和突变TAT蛋白的腺病毒和逆转录病毒载体库。 2）确定TAT两个野生型等位基因对HIV-1感染靶向的免疫细胞中宿主细胞基因表达的影响。 3）绘制负责修改宿主细胞基因表达程序的TAT域。 4）研究TAT表达过程中转录因子STAT1和IRF7的基因组规模位置，因为这些因子被HIV-1感染和TAT表达上调。 5）研究TAT使用基因组规模的位置分析实验旨在检测DNA和RNA结合的机制来研究宿主细胞基因表达的机制。