ROLES OF HOST FACTORS IN VIRAL REPLICATION COUPLED PROCESSES - 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.
