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

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

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.

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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