Cargando…

Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions

Positive-strand and double-strand RNA viruses typically compartmentalize their replication machinery in infected cells. This is thought to shield viral RNA from detection by innate immune sensors and favor RNA synthesis. The picture for the non-segmented negative-strand (NNS) RNA viruses, however, i...

Descripción completa

Detalles Bibliográficos
Autores principales: Heinrich, Bianca S., Cureton, David K., Rahmeh, Amal A., Whelan, Sean P. J.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2891829/
https://www.ncbi.nlm.nih.gov/pubmed/20585632
http://dx.doi.org/10.1371/journal.ppat.1000958
_version_ 1782182905087262720
author Heinrich, Bianca S.
Cureton, David K.
Rahmeh, Amal A.
Whelan, Sean P. J.
author_facet Heinrich, Bianca S.
Cureton, David K.
Rahmeh, Amal A.
Whelan, Sean P. J.
author_sort Heinrich, Bianca S.
collection PubMed
description Positive-strand and double-strand RNA viruses typically compartmentalize their replication machinery in infected cells. This is thought to shield viral RNA from detection by innate immune sensors and favor RNA synthesis. The picture for the non-segmented negative-strand (NNS) RNA viruses, however, is less clear. Working with vesicular stomatitis virus (VSV), a prototype of the NNS RNA viruses, we examined the location of the viral replication machinery and RNA synthesis in cells. By short-term labeling of viral RNA with 5′-bromouridine 5′-triphosphate (BrUTP), we demonstrate that primary mRNA synthesis occurs throughout the host cell cytoplasm. Protein synthesis results in the formation of inclusions that contain the viral RNA synthesis machinery and become the predominant sites of mRNA synthesis in the cell. Disruption of the microtubule network by treatment of cells with nocodazole leads to the accumulation of viral mRNA in discrete structures that decorate the surface of the inclusions. By pulse-chase analysis of the mRNA, we find that viral transcripts synthesized at the inclusions are transported away from the inclusions in a microtubule-dependent manner. Metabolic labeling of viral proteins revealed that inhibiting this transport step diminished the rate of translation. Collectively those data suggest that microtubule-dependent transport of viral mRNAs from inclusions facilitates their translation. Our experiments also show that during a VSV infection, protein synthesis is required to redirect viral RNA synthesis to intracytoplasmic inclusions. As viral RNA synthesis is initially unrestricted, we speculate that its subsequent confinement to inclusions might reflect a cellular response to infection.
format Text
id pubmed-2891829
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-28918292010-06-28 Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions Heinrich, Bianca S. Cureton, David K. Rahmeh, Amal A. Whelan, Sean P. J. PLoS Pathog Research Article Positive-strand and double-strand RNA viruses typically compartmentalize their replication machinery in infected cells. This is thought to shield viral RNA from detection by innate immune sensors and favor RNA synthesis. The picture for the non-segmented negative-strand (NNS) RNA viruses, however, is less clear. Working with vesicular stomatitis virus (VSV), a prototype of the NNS RNA viruses, we examined the location of the viral replication machinery and RNA synthesis in cells. By short-term labeling of viral RNA with 5′-bromouridine 5′-triphosphate (BrUTP), we demonstrate that primary mRNA synthesis occurs throughout the host cell cytoplasm. Protein synthesis results in the formation of inclusions that contain the viral RNA synthesis machinery and become the predominant sites of mRNA synthesis in the cell. Disruption of the microtubule network by treatment of cells with nocodazole leads to the accumulation of viral mRNA in discrete structures that decorate the surface of the inclusions. By pulse-chase analysis of the mRNA, we find that viral transcripts synthesized at the inclusions are transported away from the inclusions in a microtubule-dependent manner. Metabolic labeling of viral proteins revealed that inhibiting this transport step diminished the rate of translation. Collectively those data suggest that microtubule-dependent transport of viral mRNAs from inclusions facilitates their translation. Our experiments also show that during a VSV infection, protein synthesis is required to redirect viral RNA synthesis to intracytoplasmic inclusions. As viral RNA synthesis is initially unrestricted, we speculate that its subsequent confinement to inclusions might reflect a cellular response to infection. Public Library of Science 2010-06-24 /pmc/articles/PMC2891829/ /pubmed/20585632 http://dx.doi.org/10.1371/journal.ppat.1000958 Text en Heinrich et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Heinrich, Bianca S.
Cureton, David K.
Rahmeh, Amal A.
Whelan, Sean P. J.
Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
title Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
title_full Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
title_fullStr Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
title_full_unstemmed Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
title_short Protein Expression Redirects Vesicular Stomatitis Virus RNA Synthesis to Cytoplasmic Inclusions
title_sort protein expression redirects vesicular stomatitis virus rna synthesis to cytoplasmic inclusions
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2891829/
https://www.ncbi.nlm.nih.gov/pubmed/20585632
http://dx.doi.org/10.1371/journal.ppat.1000958
work_keys_str_mv AT heinrichbiancas proteinexpressionredirectsvesicularstomatitisvirusrnasynthesistocytoplasmicinclusions
AT curetondavidk proteinexpressionredirectsvesicularstomatitisvirusrnasynthesistocytoplasmicinclusions
AT rahmehamala proteinexpressionredirectsvesicularstomatitisvirusrnasynthesistocytoplasmicinclusions
AT whelanseanpj proteinexpressionredirectsvesicularstomatitisvirusrnasynthesistocytoplasmicinclusions