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

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 structu...

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Autores principales: Neidermyer, William J., Whelan, Sean P. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6608984/
https://www.ncbi.nlm.nih.gov/pubmed/31226162
http://dx.doi.org/10.1371/journal.ppat.1007875
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author Neidermyer, William J.
Whelan, Sean P. J.
author_facet Neidermyer, William J.
Whelan, Sean P. J.
author_sort Neidermyer, William J.
collection PubMed
description 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.
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spelling pubmed-66089842019-07-12 Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells Neidermyer, William J. Whelan, Sean P. J. PLoS Pathog Research Article 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. Public Library of Science 2019-06-21 /pmc/articles/PMC6608984/ /pubmed/31226162 http://dx.doi.org/10.1371/journal.ppat.1007875 Text en © 2019 Neidermyer, Whelan http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Neidermyer, William J.
Whelan, Sean P. J.
Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
title Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
title_full Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
title_fullStr Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
title_full_unstemmed Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
title_short Global analysis of polysome-associated mRNA in vesicular stomatitis virus infected cells
title_sort global analysis of polysome-associated mrna in vesicular stomatitis virus infected cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6608984/
https://www.ncbi.nlm.nih.gov/pubmed/31226162
http://dx.doi.org/10.1371/journal.ppat.1007875
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