Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells.


Journal

PLoS pathogens
ISSN: 1553-7374
Titre abrégé: PLoS Pathog
Pays: United States
ID NLM: 101238921

Informations de publication

Date de publication:
06 2019
Historique:
received: 22 08 2018
accepted: 27 05 2019
revised: 03 07 2019
pubmed: 22 6 2019
medline: 4 12 2019
entrez: 22 6 2019
Statut: epublish

Résumé

Infection of mammalian cells with vesicular stomatitis virus (VSV) results in the inhibition of cellular translation while viral translation proceeds efficiently. VSV RNA synthesis occurs entirely within the cytoplasm, where during transcription the viral polymerase produces 5 mRNAs that are structurally indistinct to cellular mRNAs with respect to their 5' cap-structure and 3'-polyadenylate tail. Using the global approach of massively parallel sequencing of total cytoplasmic, monosome- and polysome-associated mRNA, we interrogate the impact of VSV infection of HeLa cells on translation. Analysis of sequence reads in the different fractions shows >60% of total cytoplasmic and polysome-associated reads map to the 5 viral genes by 6 hours post-infection, a time point at which robust host cell translational shut-off is observed. Consistent with an overwhelming abundance of viral mRNA in the polysome fraction, the reads mapping to cellular genes were reduced. The cellular mRNAs that remain most polysome-associated following infection had longer half-lives, were typically larger, and were more AU rich, features that are shared with the viral mRNAs. Several of those mRNAs encode proteins known to positively affect viral replication, and using chemical inhibition and siRNA depletion we confirm that the host chaperone heat shock protein 90 (hsp90) and eukaryotic translation initiation factor 3A (eIF3A)-encoded by 2 such mRNAs-support viral replication. Correspondingly, regulated in development and DNA damage 1 (Redd1) encoded by a host mRNA with reduced polysome association inhibits viral infection. These data underscore the importance of viral mRNA abundance in the shut-off of host translation in VSV infected cells and link the differential translatability of some cellular mRNAs with pro- or antiviral function.

Identifiants

pubmed: 31226162
doi: 10.1371/journal.ppat.1007875
pii: PPATHOGENS-D-18-01660
pmc: PMC6608984
doi:

Substances chimiques

RNA, Messenger 0

Types de publication

Journal Article Research Support, N.I.H., Extramural

Langues

eng

Sous-ensembles de citation

IM

Pagination

e1007875

Subventions

Organisme : NIAID NIH HHS
ID : R37 AI059371
Pays : United States

Déclaration de conflit d'intérêts

The authors have declared that no competing interests exist.

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Auteurs

William J Neidermyer (WJ)

Department of Microbiology & Immunobiology, Program in Virology, Harvard Medical School, Boston, Massachusetts, United States of America.

Sean P J Whelan (SPJ)

Department of Microbiology & Immunobiology, Program in Virology, Harvard Medical School, Boston, Massachusetts, United States of America.

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