Molecular Genetics of HSV Reactivation

描述：（由申请人提供）：疱疹病毒普遍存在， 造成了大量的人类死亡和痛苦， 初始感染和（甚至更多）复发。此外，随着越来越多 免疫抑制性疾病的发生， 具有临床意义的初始和潜伏疱疹病毒感染的频率 起来。因此，该项目的长期目标是获得 疱疹病毒编码基因参与潜伏期的功能理解 和重新激活-知识是基本的理性设计， 介入治疗我们已经确定了基因组中的一个区域， "再激活关键区"（RCR）是肾上腺素诱导的细胞凋亡所必需的。 重新激活本提案的主要重点是描述 单纯疱疹病毒1型rcr中的功能性作用序列元件 (HSV-1）基因组在感觉神经节中潜伏感染的再激活中发挥作用 神经元 本研究的主要假设是HSV rcr促进 通过调节潜在基因组的基因表达， 在再激活期间启动急性基因表达。使用分子 涉及专门工程病毒重组的遗传方法，我们将 遵循HSV-1再激活的转录和复制过程， 兔角膜-肾上腺素模型将病毒遗传学与 动物具体而言，拟议的研究将测试三种可能性， 难民救济委员会促进恢复活动的机制： 作为转录的调节剂，其通过以下方式促进再激活： 调节LAT、ICPO和/或ICP 4的表达，B）rcr是 甲基化和/或在潜伏期调节转录的细胞因子 和再活化，以及c）rcr作为非编码功能性RNA， 通过剂量补偿样机制调节基因表达。这三 这些机制并不相互排斥，它们之间可能存在相互作用。 这些调节元件对于维持和重新激活 从latency。这项工作将导致确定病毒的目标， 宿主因素传达压力刺激导致再激活。

DESCRIPTION:(provided by applicant): Herpesviruses are ubiquitous and are responsible for significant human mortality and suffering both in terms of initial infections and (even more so) recurrences. In addition, with increasing occurrences of immunosuppressive disorders, a corresponding increase in the frequency of clinically significant initial and latent herpesvirus infections arise. Therefore, the long-term objective of this project is to gain a functional understanding of the herpesvinis-encoded genes involved in latency and reactivation - knowledge that is fundamental to the rational design of interventive therapies. We have identified a region of the genome termed the "reactivation critical region" (rcr) that is required for epinephrine-induced reactivation. The primary focus of this proposal is to characterize the functional role sequence elements in the rcr of the Herpes simplex virus type 1 (HSV- 1) genome play in reactivation of infections latent in sensory ganglia neurons. The overriding hypothesis of this study is that the HSV rcr facilitates reactivation by regulating gene expression from the latent genome allowing the initiation of acute gene expression during reactivation. Using a molecular genetic approach involving specifically engineered viral recombinants, we will follow the transcriptional and replicative processes of HSV- 1 reactivation in the rabbit cornea-epinephrine model to link viral genetics with processes in animals. Specifically, the proposed studies will test three potential mechanisms by which the rcr functions to facilitate reactivation: a) the rcr acts as a modulator of transcription which facilitates reactivation by regulating the expression of LAT, ICPO and/or ICP4, b) the rcr is a target of methylation and/or cellular factors that regulate transcription during latency and reactivation, and c) the rcr acts as a non-coding functional RNA that regulates gene expression via a dosage compensation-like mechanism. These three mechanisms are not mutually exclusive and it is likely an interplay between these regulatory elements is necessary for the maintenance of and reactivation from latency. This work will lead to the identification of viral target(s) of host factors that communicate stress stimuli leading to reactivation.