Journal
JOURNAL OF VIROLOGY
Volume 95, Issue 3, Pages -Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.01399-20
Keywords
4sU-seq; herpes simplex virus 1; RNA degradation; RNA-seq; chromatin-associated RNA; proteomics; transcriptional regulation; virion host shutoff protein
Categories
Funding
- Deutsche Forschungsgemeinschaft (DFG) [D01275/2-1, D01275/6-1, FR2938/7-1, FR2938/9-1]
- Medical Research Council [CSF G1002523, MR/P008801/1]
- Wellcome Trust [PRF 210688/Z/18/Z]
- NHS Blood and Transplant [WP11-05, WPA15-02]
- European Research Council [ERC-2016-CoG 721016]
- NIHR Cambridge BRC
- Alexander von Humboldt Foundation
- German Federal Foreign Office
- MRC [MR/P008801/1] Funding Source: UKRI
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During the early stages of HSV-1 infection, the vhs protein cleaves host and viral mRNAs and decreases host transcriptional activity, leading to changes in RNA metabolism. While alterations in total RNA levels were mainly influenced by these global effects, gene-specific transcriptional changes were observed in chromatin-associated RNA.
Herpes simplex virus 1 (HSV-1) induces a profound host shutoff during lytic infection. The virion host shutoff (vhs) protein plays a key role in this process by efficiently cleaving host and viral mRNAs. Furthermore, the onset of viral DNA replication is accompanied by a rapid decline in host transcriptional activity. To dissect relative contributions of both mechanisms and elucidate gene-specific host transcriptional responses throughout the first 8 h of lytic HSV-1 infection, we used transcriptome sequencing of total, newly transcribed (4sU-labeled) and chromatin-associated RNA in wild-type (WT) and Delta vhs mutant infection of primary human fibroblasts. Following virus entry, vhs activity rapidly plateaued at an elimination rate of around 30% of cellular mRNAs per hour until 8 h postinfection (p.i.). In parallel, host transcriptional activity dropped to 10 to 20%. While the combined effects of both phenomena dominated infection-induced changes in total RNA, extensive gene-specific transcriptional regulation was observable in chromatin-associated RNA and was surprisingly concordant between WT and Delta vhs infections. Both induced strong transcriptional upregulation of a small subset of genes that were poorly expressed prior to infection but already primed by H3K4me3 histone marks at their promoters. Most interestingly, analysis of chromatin-associated RNA revealed vhsnuclease-activity-dependent transcriptional downregulation of at least 150 cellular genes, in particular of many integrin adhesome and extracellular matrix components. This was accompanied by a vhs-dependent reduction in protein levels by 8 h p.i. for many of these genes. In summary, our study provides a comprehensive picture of the molecular mechanisms that govern cellular RNA metabolism during the first 8 h of lytic HSV-1 infection. IMPORTANCE The HSV-1 virion host shutoff (vhs) protein efficiently cleaves both host and viral mRNAs in a translation-dependent manner. In this study, we model and quantify changes in vhs activity, as well as virus-induced global loss of host transcriptional activity, during productive HSV-1 infection. In general, HSV-1-induced alterations in total RNA levels were dominated by these two global effects. In contrast, chromatin-associated RNA depicted gene-specific transcriptional changes. This revealed highly concordant transcriptional changes in WT and Delta vhs infections, confirmed DUX4 as a key transcriptional regulator in HSV-1 infection, and identified vhsdependent transcriptional downregulation of the integrin adhesome and extracellular matrix components. The latter explained seemingly gene-specific effects previously attributed to vhs-mediated mRNA degradation and resulted in a concordant loss in protein levels by 8 h p.i. for many of the respective genes.
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