ICP22 of Herpes Simplex Virus 1 decreases RNA Polymerase Processivity

These data files consist of Illumina next seq 500 sequencing data from precision nuclear run-on and global nuclear run-on experiments conducted with human epithelial Hep2 cells that have been infected with human herpes simplex virus-1 (F) strain mutants. The results of these studies suggest that ICP22 is necessary for reducing Pol II processivity on the viral genome which results in its maintenance on the viral genome over the course of infection. While human reads continued to decrease over the time course of infection in the repair virus, this did not occur in the ICP22 deletion virus where over time the number of human reads increased over the time course of infection rather than decreasing. The effects of ICP22 deletion were separable from inhibiting HSV-1 DNA replication to the extent that ICP22 deletion resulted in a decrease in Pol levels only at 6 hpi and not at 3 hpi as was observed in the mutant PRO-seq and GRO-seq were conducted on cells that had been infected with two different HSV-1 (F) strain viruses. Infection occurred for either 3 or 6 hours before collection of nuclei for PRO/GRO-seq processing, and the mutant viruses used were a bacterial artificial choromosome derived virus with the ICP22 open reading frame deleted or bearing a genetically restored virus. These viruses were generated by Maruzuru et. al., 2013, Roles of p53 in Herpes Simplex Virus 1 Replication, Journal of Virology. After nuclei extraction, and before the run-on reactions, extracted Drosophila melanogaster nuclei were spiked into infected nuclei solution at a ratio of 1 Drosophila melanogaster nuclei for every 1000 infected Homo sapiens nuclei. Sequencing libraries were prepared from the biotinylated RNA's in the PRO-seq or the Br-UTP RNAs produced during either 5 min or 30 min of GRO-seq run-on for each replicate of each virus at each time point post infection.