Regulation of HIV Transcription Elongation

HIV转录延伸的调控

• 批准号: 7876845
• 负责人: Andrew J Henderson
• 金额: $ 41.75万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-19 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7876845
• 关键词: Binding; Biochemical; Cell Line; Cells; Chromatin; Complex; Coupled; Data; Drug resistance; Event; Genes; Genetic Transcription; Goals; HIV; HIV Infections; Histone Acetylation; Histone Deacetylase; Histones; In Vitro; Maintenance; Mass Spectrum Analysis; Molecular; Nucleosomes; Polymerase; Population; Positioning Attribute; Positive Transcriptional Elongation Factor B; Post-Translational Protein Processing; Printing; Process; Proviruses; RNA Polymerase II; Regulation; Reporting; Site; Small Interfering RNA; Suggestion; System; Transcription Elongation; Transcription Initiation; Viral; Viral Physiology; Virus; base; cellular targeting; chromatin immunoprecipitation; chromatin remodeling; foot; histone modification; in vivo; inhibitor/antagonist; insight; negative elongation factor; novel; permanganate; premature; public health relevance; purge; resistant strain; termination factor; transcription factor; transcription termination

DESCRIPTION (provided by applicant): Studies of the molecular events that establish and maintain latency have been hampered by the rarity and inaccessibility of latently infected cells. HIV provirus is regulated at the level of transcription and involves changes in transcription initiation, chromatin organization and elongation. In particular, transcription elongation has been demonstrated to be a limiting step for HIV expression and overcoming this block is the primary activity of the viral factor Tat. We hypothesize that cellular factors that regulate transcription elongation and premature transcription termination have a direct impact on HIV transcription, including extinguishing HIV transcription and establishing latency. Preliminary data show that Pol II processivity is a check point for HIV transcription in the absence of Tat and that two factors, NELF and Pcf11, negatively regulate HIV transcription elongation. Using a variety of biochemical approaches, including chromatin immunoprecipitation, Pol II foot printing and in vivo and in vitro transcription systems, we propose to determine the biochemical mechanisms and characterize the cellular factors that negatively regulate HIV transcription elongation. We expect that these studies will provide new insights into the establishment, maintenance and reversal of HIV latency. Understanding the regulation of HIV transcription elongation will provide novel cellular targets for controlling and purging HIV in different cellular reservoirs. PUBLIC HEALTH RELEVANCE: Successfully eradicating HIV infection will require understanding how cellular reservoirs that poorly express virus are established and maintained as well as determining whether these populations can be purged of HIV. This proposal focuses on the biochemical mechanisms that repress HIV transcription, in order to gain insights into factors that regulate HIV latency with long term goals of identifying novel targets for controlling HIV expression in different cells.

描述（由申请人提供）：对建立和维持潜伏期的分子事件的研究受到了潜在感染细胞的稀有性和无法访问的阻碍。 HIV病毒受到转录水平的调节，涉及转录起始，染色质组织和伸长率的变化。特别是，已证明转录伸长是HIV表达的限制步骤，克服该障碍是病毒因子TAT的主要活性。我们假设调节转录伸长和过早转录终止的细胞因子对HIV转录有直接影响，包括熄灭HIV转录和确定潜伏期。初步数据表明，在没有TAT的情况下，POL II加工性是HIV转录的一个检查点，NELF和PCF11的两个因素对HIV转录伸长延伸。使用多种生化方法，包括染色质免疫沉淀，POL II脚印以及体内和体外转录系统，我们建议确定生化机制，并表征对HIV转录延长的细胞因子的表征。我们预计这些研究将为艾滋病毒潜伏期的建立，维护和逆转提供新的见解。了解HIV转录伸长的调节将为控制和清除不同细胞储层中的HIV提供新颖的细胞靶标。公共卫生相关性：成功消除艾滋病毒感染将需要了解如何确定和维持较差病毒的细胞储层以及确定是否可以清除艾滋病毒的人群。该提案的重点是抑制HIV转录的生化机制，以便深入了解调节HIV潜伏期的因素，这些因素具有长期目标，即确定了控制不同细胞中HIV表达的新目标。