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Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA

Viruses that generate capped RNA lacking 2′O methylation on the first ribose are severely affected by the antiviral activity of Type I interferons. We used proteome-wide affinity purification coupled to mass spectrometry to identify human and mouse proteins specifically binding to capped RNA with di...

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Autores principales: Habjan, Matthias, Hubel, Philipp, Lacerda, Livia, Benda, Christian, Holze, Cathleen, Eberl, Christian H., Mann, Angelika, Kindler, Eveline, Gil-Cruz, Cristina, Ziebuhr, John, Thiel, Volker, Pichlmair, Andreas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3789756/
https://www.ncbi.nlm.nih.gov/pubmed/24098121
http://dx.doi.org/10.1371/journal.ppat.1003663
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author Habjan, Matthias
Hubel, Philipp
Lacerda, Livia
Benda, Christian
Holze, Cathleen
Eberl, Christian H.
Mann, Angelika
Kindler, Eveline
Gil-Cruz, Cristina
Ziebuhr, John
Thiel, Volker
Pichlmair, Andreas
author_facet Habjan, Matthias
Hubel, Philipp
Lacerda, Livia
Benda, Christian
Holze, Cathleen
Eberl, Christian H.
Mann, Angelika
Kindler, Eveline
Gil-Cruz, Cristina
Ziebuhr, John
Thiel, Volker
Pichlmair, Andreas
author_sort Habjan, Matthias
collection PubMed
description Viruses that generate capped RNA lacking 2′O methylation on the first ribose are severely affected by the antiviral activity of Type I interferons. We used proteome-wide affinity purification coupled to mass spectrometry to identify human and mouse proteins specifically binding to capped RNA with different methylation states. This analysis, complemented with functional validation experiments, revealed that IFIT1 is the sole interferon-induced protein displaying higher affinity for unmethylated than for methylated capped RNA. IFIT1 tethers a species-specific protein complex consisting of other IFITs to RNA. Pulsed stable isotope labelling with amino acids in cell culture coupled to mass spectrometry as well as in vitro competition assays indicate that IFIT1 sequesters 2′O-unmethylated capped RNA and thereby impairs binding of eukaryotic translation initiation factors to 2′O-unmethylated RNA template, which results in inhibition of translation. The specificity of IFIT1 for 2′O-unmethylated RNA serves as potent antiviral mechanism against viruses lacking 2′O-methyltransferase activity and at the same time allows unperturbed progression of the antiviral program in infected cells.
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spelling pubmed-37897562013-10-04 Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA Habjan, Matthias Hubel, Philipp Lacerda, Livia Benda, Christian Holze, Cathleen Eberl, Christian H. Mann, Angelika Kindler, Eveline Gil-Cruz, Cristina Ziebuhr, John Thiel, Volker Pichlmair, Andreas PLoS Pathog Research Article Viruses that generate capped RNA lacking 2′O methylation on the first ribose are severely affected by the antiviral activity of Type I interferons. We used proteome-wide affinity purification coupled to mass spectrometry to identify human and mouse proteins specifically binding to capped RNA with different methylation states. This analysis, complemented with functional validation experiments, revealed that IFIT1 is the sole interferon-induced protein displaying higher affinity for unmethylated than for methylated capped RNA. IFIT1 tethers a species-specific protein complex consisting of other IFITs to RNA. Pulsed stable isotope labelling with amino acids in cell culture coupled to mass spectrometry as well as in vitro competition assays indicate that IFIT1 sequesters 2′O-unmethylated capped RNA and thereby impairs binding of eukaryotic translation initiation factors to 2′O-unmethylated RNA template, which results in inhibition of translation. The specificity of IFIT1 for 2′O-unmethylated RNA serves as potent antiviral mechanism against viruses lacking 2′O-methyltransferase activity and at the same time allows unperturbed progression of the antiviral program in infected cells. Public Library of Science 2013-10-03 /pmc/articles/PMC3789756/ /pubmed/24098121 http://dx.doi.org/10.1371/journal.ppat.1003663 Text en © 2013 Habjan 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
Habjan, Matthias
Hubel, Philipp
Lacerda, Livia
Benda, Christian
Holze, Cathleen
Eberl, Christian H.
Mann, Angelika
Kindler, Eveline
Gil-Cruz, Cristina
Ziebuhr, John
Thiel, Volker
Pichlmair, Andreas
Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA
title Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA
title_full Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA
title_fullStr Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA
title_full_unstemmed Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA
title_short Sequestration by IFIT1 Impairs Translation of 2′O-unmethylated Capped RNA
title_sort sequestration by ifit1 impairs translation of 2′o-unmethylated capped rna
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3789756/
https://www.ncbi.nlm.nih.gov/pubmed/24098121
http://dx.doi.org/10.1371/journal.ppat.1003663
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