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Ribose 2′-O-methylation provides a molecular signature for the distinction of self and non-self mRNA dependent on the RNA sensor Mda5

Nature Immunology volume 12, pages 137–143 (2011)Cite this article

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Abstract

The 5′ cap structures of higher eukaryote mRNAs have ribose 2′-O-methylation. Likewise, many viruses that replicate in the cytoplasm of eukaryotes have evolved 2′-O-methyltransferases to autonomously modify their mRNAs. However, a defined biological role for 2′-O-methylation of mRNA remains elusive. Here we show that 2′-O-methylation of viral mRNA was critically involved in subverting the induction of type I interferon. We demonstrate that human and mouse coronavirus mutants lacking 2′-O-methyltransferase activity induced higher expression of type I interferon and were highly sensitive to type I interferon. Notably, the induction of type I interferon by viruses deficient in 2′-O-methyltransferase was dependent on the cytoplasmic RNA sensor Mda5. This link between Mda5-mediated sensing of viral RNA and 2′-O-methylation of mRNA suggests that RNA modifications such as 2′-O-methylation provide a molecular signature for the discrimination of self and non-self mRNA.

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Figure 1: Conservation of viral 2′-O-methyltransferases.
Figure 2: The HCoV 2′-O-methyltransferase mutant has altered replication kinetics and induction of and sensitivity to type I interferon.
Figure 3: MHV 2′-O-methyltransferase mutants induce IFN-β in an Mda5-dependent manner.
Figure 4: MHV 2′-O-methyltransferase mutants induce the nuclear localization of IRF3 in an Mda5-dependent manner.
Figure 5: Differences in the effect of type I interferon on the replication of MHV 2′-O-methyltransferase mutants.
Figure 6: Restoration of MHV-D130A replication in IFIT1-deficient macrophages.
Figure 7: Deficiency in 2′-O-methyltransferase affects the recognition of virus by the innate immune system in vivo.

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Acknowledgements

We thank V. Lohmann (University of Heidelberg) for Huh-7 cells; G.L. Smith (Imperial College, London) for D980R cells; S.G. Sawicki (Medical University of Ohio) for 17Cl1 cells; M. Pelegrin (Institut de Génétique Moléculaire de Montpellier) for LL171 cells; L. Onder for assistance with fluorescence microscopy; and R. de Giuli, B. Schelle and N. Karl for technical assistance. This study was supported by the Swiss National Science Foundation, the European Commission (TOLERAGE), the Novartis Foundation for Biomedical Research, Switzerland, the German Ministry of Education and Research (V.T.), the Austrian Science Fund (FWF J3044 to M.H.), Deutsche Forschungsgemeinschaft (J.Z.), the National Institutes of Health (AI060915 and AI085089 to S.C.B.; U54 AI081680 (Pacific Northwest Regional Center of Excellence for Biodefense and Emerging Infectious Diseases Research to M.S.D), the Medical Research Council (B.W.N.) and the Wellcome Trust (S.G.S.).

Author information

Author notes

  1. Roland Züst

    Present address: Present address: Singapore Immunology Network, Agency for Science, Technology and Research, Singapore.,

  2. Roland Züst and Luisa Cervantes-Barragan: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Immunobiology, Kantonal Hospital St. Gallen, St. Gallen, Switzerland

    Roland Züst, Luisa Cervantes-Barragan, Matthias Habjan, Reinhard Maier, Burkhard Ludewig & Volker Thiel

  2. School of Biological Sciences, University of Reading, Reading, UK

    Benjamin W Neuman

  3. Centre for Infection and Immunity, Queen's University Belfast, Belfast, UK

    John Ziebuhr

  4. Institute of Medical Virology, Justus Liebig University Giessen, Giessen, Germany

    John Ziebuhr

  5. Department of Medicine, Department of Molecular Microbiology, and Department of Pathology & Immunology, Washington University School of Medicine, St. Louis, Missouri, USA

    Kristy J Szretter & Michael S Diamond

  6. Department of Microbiology and Immunology, Loyola University Stritch School of Medicine, Maywood, Illinois, USA

    Susan C Baker

  7. Institute for Clinical Chemistry and Pharmacology, University Hospital, University of Bonn, Bonn, Germany

    Winfried Barchet

  8. Department of Cellular and Molecular Medicine, School of Medical and Veterinary Sciences, University of Bristol, Bristol, UK

    Stuart G Siddell

  9. Vetsuisse Faculty, University of Zürich, Zürich, Switzerland

    Burkhard Ludewig & Volker Thiel

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Contributions

R.Z., L.C.-B., M.H., R.M. and K.J.S. did most of the experiments; B.W.N. did phylogenetic analyses; B.W.N. and S.C.B. did electron microscopy; J.Z., S.C.B., W.B., M.S.D., S.G.S. and B.L. contributed reagents and expertise; and S.G.S., B.W.N., B.L. and V.T. conceived of and designed the project and wrote and edited the manuscript.

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Correspondence to Volker Thiel.

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Züst, R., Cervantes-Barragan, L., Habjan, M. et al. Ribose 2′-O-methylation provides a molecular signature for the distinction of self and non-self mRNA dependent on the RNA sensor Mda5. Nat Immunol 12, 137–143 (2011). https://doi.org/10.1038/ni.1979

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