ROLES OF HOST FACTORS IN VIRAL REPLICATION COUPLED PROCESSES

单纯疱疹病毒是一种流行的病原体，感染了大多数人。病毒DNA复制是病毒生命周期的一个重要步骤，可以作为抗病毒治疗的目标。疱疹病毒的DNA复制与其他病毒过程耦合，包括转录、DNA重组和DNA修复。然而，在我们目前对这些过程在病毒DNA上协调和调节的分子机制的知识中存在关键空白。最近发展的蛋白质组学方法研究蛋白质与复制叉的关联，使人们能够深入研究细胞内的这一基本过程。除了病毒复制机制外，选择宿主染色质重塑，转录，DNA修饰和DNA修复因子与HSV-1复制叉相关。我们的长期目标是揭示HSV-1如何破坏宿主细胞因子来复制病毒DNA，以协调病毒DNA合成与DNA损伤反应途径，DNA修复和体内转录。在提出的目标中，蛋白质组学和全基因组分析将用于1)研究将病毒复制机制与细胞DNA修复途径耦合的机制，2)确定细胞因子如何介导复制耦合开关以激活晚期基因转录，以及3)剖析复制叉相关因子如何调节病毒DNA拓扑结构和基因组完整性。创新的方法将被用来定义宿主蛋白在病毒DNA复制过程中的功能。病毒修饰细胞因子如何改变细胞DNA复制和修复也将被研究。在病毒DNA复制过程中被利用的宿主蛋白可以被抗病毒治疗靶向。此外，深入了解病毒复制如何与DNA损伤反应、病毒染色质和转录调控协调，将有助于开发有效的基于HSV-1的基因治疗方法和溶瘤载体。

Herpes simplex virus is a prevalent pathogen that infects the majority of the human population. Viral DNA replication is an essential step in the virus life cycle and can be targeted by antiviral treatments. Herpesviral DNA replication is coupled to other viral processes including transcription, DNA recombination, and DNA repair. However, there are critical gaps in our current knowledge of the molecular mechanisms by which these processes are coordinated and regulated on the viral DNA. The recent development of proteomic approaches to study protein association with replication forks has enabled an in depth look at this fundamental process within cells. In addition to the viral replication machinery, select host chromatin remodeling, transcription, DNA modifying, and DNA repair factors associate with HSV-1 replication forks. Our long-term goal is to uncover how HSV-1 subverts host cellular factors to replicating viral DNA to coordinate viral DNA synthesis with DNA damage response pathways, DNA repair, and transcription in vivo. In the proposed aims, proteomic and genome-wide assays will be used to 1) investigate mechanisms that couple the viral replication machinery to cellular DNA repair pathways, 2) determine how cellular factors mediate the replication coupled switch to activate late gene transcription, and 3) dissect how replication fork associated factors regulate viral DNA topology and genome integrity. Innovative approaches will be employed to define the functions of host proteins during viral DNA replication. How viral modification of cellular factors alters cellular DNA replication and repair will also be investigated. Host proteins that are exploited during viral DNA replication can be targeted by antiviral therapies. Furthermore, insight into how viral replication is coordinated with the DNA damage response, viral chromatin, and transcription regulation will aid in the development of effective HSV-1 based gene therapy approaches and oncolytic vectors.