ROLES OF HOST FACTORS IN VIRAL REPLICATION COUPLED PROCESSES

单纯疱疹病毒是一种流行的病原体，感染大多数人类人口。病毒DNA复制是病毒生命周期中的一个重要步骤，可以作为抗病毒治疗的靶点。疱疹病毒DNA复制与其他病毒过程相耦合，包括转录、DNA重组和DNA修复。然而，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了解存在着严重的空白。最近蛋白质组学方法的发展，以研究蛋白质与复制叉子的关系，使我们能够深入研究细胞内的这一基本过程。除病毒复制机制外，选择与HSV-1复制叉相关的宿主染色质重塑、转录、DNA修饰和DNA修复因子。我们的长期目标是揭示HSV-1如何颠覆宿主细胞因子来复制病毒DNA，以协调病毒DNA的合成与DNA损伤反应途径、DNA修复和体内转录。在拟议的AIMS中，蛋白质组学和全基因组分析将用于1)研究病毒复制机制与细胞DNA修复途径的耦合机制，2)确定细胞因素如何介导复制耦合开关以激活晚期基因转录，以及3)剖析复制分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。病毒对细胞因子的修饰如何改变细胞的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成为抗病毒治疗的靶点。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相协调，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.