ROLE OF CHROMATIN IN HERPES SIMPLEX VIRUS TYPE 1 (HSV-1) GENE REGULATION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们的实验室研究HSV-1在裂解和潜伏感染期间的分子和细胞生物学，包括基因沉默和再激活的机制。我们研究了TR和T3在体外HSV-1基因表达中的作用，在确定了HSV-1 TK和LAT启动子中的甲状腺激素受体（TR）反应元件（TREs）后。我们证明，配体TR抑制TK启动子活性的转染和感染的神经元细胞。我们的数据还表明，配体TRs激活LAT转录，但抑制ICP 0转录。TRs独立于T3被募集到这些启动子，并且在激素存在下，超乙酰化组蛋白H4与这些启动子相关。此外，在TR和T3存在下，各种染色质修饰复合物如CTCF和BRG 1复合物被募集到这些启动子。此外，T3处理的N2 aTR？细胞在低moi下抑制TK表达和病毒排出，但激素的提取增强了TK表达，从而增加了感染性病毒的释放。这些结果首次表明T3可以调节HSV-1感染，并可能在体内病毒潜伏和再激活中起作用。使用动物模型的研究正在进行中。 我们还研究了我们与El Sayed博士合作开发的候选抗病毒药物。结果表明，生物碱Manzamine A在1 μ M时对角膜细胞系中的病毒感染具有显著的抑制作用。这些结果表明，它可能有很大的潜力，以防止HSV-1诱导的眼部感染。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our laboratory studies the molecular and cellular biology of HSV-1 during lytic and latent infections including the mechanisms of gene silencing and reactivation. We investigated the roles of TR and T3 on HSV-1 gene expression in vitro after identifying the thyroid hormone receptor (TR) responsive elements (TREs) in the promoters of HSV-1 TK and LAT. We demonstrated that liganded TR repressed TK promoter activity by transfection and infection of neuronal cells. Our data also showed that the liganded TRs activated LAT transcription but repressed ICP0 transcription. TRs were recruited to these promoters independently of T3 and hyperacetylated histones H4 was associated with these promoters in the presence of hormone. In addition, various chromatin modifying complexes such as CTCF and BRG1 complexes were recruited to these promoters in the presence of TR and T3. Furthermore, T3-treated N2aTR¿ cells were suppressive to TK expression and virus egress at low moi but the abstraction of hormone enhanced the TK expression and thus increased the release of infectious viruses. These results, for the first time, suggested that T3 could regulate the HSV-1 infections and may have a role in viral latency and reactivation in vivo. Studies using animal models are underway. We also investigate a candidate antiviral drugs that we have developed in collaboration with Dr. El Sayed. The results showed that an alkaloid Manzamine A exhibits significant inhibitory effect on viral infection in a corneal cell line at 1¿M. These results suggest that it may have great potential to prevent HSV-1 induced eye infection.