Abstract
RNA polymerase III (Pol III) transcribes noncoding RNA, including transfer RNA (tRNA), and is commonly targeted during cancer and viral infection. We find that Herpes Simplex Virus-1 (HSV-1) stimulates tRNA expression 10-fold. Perturbation of host tRNA synthesis requires nuclear viral entry, but not synthesis of specific viral transcripts. tRNA with a specific codon bias were not targeted—rather increased transcription was observed from euchromatic, actively transcribed loci. tRNA upregulation is linked to unique crosstalk between the Pol II and III transcriptional machinery. While viral infection results in depletion of Pol II on host mRNA promoters, we find that Pol II binding to tRNA loci increases. Finally, we report Pol III and associated factors bind the viral genome, which suggests a previously unrecognized role in HSV-1 gene expression. These findings provide insight into mechanisms by which HSV-1 alters the host nuclear environment, shifting key processes in favor of the pathogen.
Highlights
Background strainKOS KOS KOS KOS KOS KOS KOS KOS F F F KOS N/A N/ATable 1 contains a detailed list of all viruses used in this study. n199, R3616, FΔICP47, and KOS were prepared and titered in Vero cells
ΔICP4 infection phenocopied transfer RNA (tRNA) transcriptional changes in wild-type infection, indicating that synthesis of E and L viral proteins, nascent viral genomes, or viral progeny was not required for alteration of the polymerase III (Pol III) transcriptional landscape
We found that Herpes Simplex Virus-1 (HSV-1) targets and upregulates tRNA, another group of Pol III type II transcripts
Summary
Background strainKOS KOS KOS KOS KOS KOS KOS KOS F F F KOS N/A N/ATable 1 contains a detailed list of all viruses used in this study. n199, R3616, FΔICP47, and KOS were prepared and titered in Vero cells. KOS KOS KOS KOS KOS KOS KOS KOS F F F KOS N/A N/A. N199, R3616, FΔICP47, and KOS were prepared and titered in Vero cells. Other virus stocks were prepared and titered in the following Vero-based complementing cell lines: E5 (ICP4 + , n12, d120-complementing41); E11 (ICP4/ICP27+, 5dl1.2, d92complementing42); F06 (ICP4/ICP27/ICP0 +, d109-complementing43); U2OS (n212, d99-complementing), POLB3 (hp66-complementing[44]). We thank Don Coen (hp[66], POLB3), David A. Leib (R3616), and Anthony St. Leger (F-ΔICP47, FΔICP47-R) for their kind gift of viruses and/or cells. A list of all antibodies and the amount used per technique is located in Supplementary Table 1
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