Molecular Analysis of HSV-1 Reactivation from Latency

HSV-1 潜伏期重新激活的分子分析

• 批准号: 7014586
• 负责人: Nancy M. Sawtell
• 金额: $ 28.43万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-01-01 至 2008-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7014586
• 关键词: DNA binding protein; gene dosage; gene induction /repression; genetic promoter element; genetic transcription; genetic translation; herpes simplex virus 1; laboratory mouse; latent virus infection; molecular genetics; neurons; virus genetics; virus infection mechanism; virus protein; virus replication

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) infection continues at epidemic levels in the global human population with serious health consequences. Intervention of the reactivation of HSV from the latent state is a critical component of efforts to control this epidemic. The long-term goal of the proposed research is to acquire knowledge of the molecular mechanisms controlling the transition from latent to lytic phase transcription for use in the rational design of intervention strategies. It is widely accepted that the viral/host cell interactions initiating the viral lytic cascade during primary infection are distinct from those initiating the lytic cascade from the latent viral genome. This notion is rooted in the fact that the alpha gene transinducing factor (VP16) is brought in with the virion as a protein and not made until late in the lytic cycle when it is incorporated as protein into the virion. Thus this important viral transactivator is thought to be absent during latency as well as during activation of transcription from the latent viral genome. This presents the necessity to identify a viral/host transcriptional activation interface other than the well-defined VP16/Octl/HCF/taatgarat complex. Although the IE gene ICP0 has been viewed as the likely candidate to "replace" VP16 in initiating the lytic cycle, and studies of ICP0 function have revealed much about the role of this protein in modifying the host cell environment, there remains no evidence that ICP0 initiates entry into the lytic cycle. We have generated compelling data, both phenotypic and biochemical that marks VP16 as a critical player in the initiation of reactivation. This key observation calls for a focused examination of the mechanism of upregulation of viral IE gene transcription from latency. The following specific aims will guide us through a systematic dissection of (1) the role of VP16 during the initiation of reactivation; (2) the VP16 promoter elements required for its stress driven upregulation; and (3) the role of viral genome structure in the regulation of transcriptional silencing and initiation of reactivation. The development and refinement of methodologies to facilitate routine examination and quantification of the events occurring in individual neurons in the ganglia have been fundamental to our progress. These analytical tools together with precision engineering of the viral genome and advances in the analysis of chromatin structure will be utilized for these studies.

描述（由申请人提供）：单纯疱疹病毒（HSV）感染在全球人口的流行水平上继续存在，其健康后果严重。 HSV从潜在状态重新激活的干预是控制这种流行病的努力的关键组成部分。拟议研究的长期目标是获取控制从潜在到裂解相转录过渡的分子机制，以在干预策略的合理设计中使用。普遍认为，在初次感染期间启动病毒裂解的病毒/宿主相互作用与从潜在病毒基因组发起裂解级联的裂解层不同。该概念植根于以下事实：α基因透射诱导因子（VP16）以蛋白质作为蛋白质引入，而直到在裂解周期后期将其作为蛋白掺入蛋白质中。因此，这种重要的病毒式反式激活剂被认为在潜伏期以及从潜在病毒基因组的转录激活过程中不存在。这提出了除了定义明确的VP16/OCTL/HCF/TAATGARAT复合物以外的病毒/宿主转录激活界面的必要性。尽管IE基因ICP0被视为可能在启动裂解周期中“替换” VP16的候选者，而对ICP0功能的研究已经揭示了该蛋白质在修改宿主细胞环境中的作用，但仍然没有证据表明ICP0启动进入裂解周期。我们已经生成了引人注目的数据，既可以将VP16标记为启动重新激活的关键参与者。该关键观察要求将重点检查病毒IE基因转录的上调机制进行重点检查。以下具体目的将指导我们通过（1）VP16在启动重新激活过程中的作用； （2）其应力驱动上调所需的VP16启动子元素； （3）病毒基因组结构在转录沉默调节和重新激活的开始中的作用。促进神经节中各个神经元中发生的事件的常规检查和定量方法的方法的发展和完善对我们的进步至关重要。这些分析工具以及病毒基因组的精确工程以及染色质结构的进步将用于这些研究。