2′-O methylation of the viral mRNA cap evades host restriction by IFIT family members

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Cellular messenger RNA (mRNA) of higher eukaryotes and many viral RNAs are methylated at the N-7 and 2′-O positions of the 5′ guanosine cap by specific nuclear and cytoplasmic methyltransferases (MTases), respectively. Whereas N-7 methylation is essential for RNA translation and stability1, the function of 2′-O methylation has remained uncertain since its discovery 35 years ago2,3,4. Here we show that a West Nile virus (WNV) mutant (E218A) that lacks 2′-O MTase activity was attenuated in wild-type primary cells and mice but was pathogenic in the absence of type I interferon (IFN) signalling. 2′-O methylation of viral RNA did not affect IFN induction in WNV-infected fibroblasts but instead modulated the antiviral effects of IFN-induced proteins with tetratricopeptide repeats (IFIT), which are interferon-stimulated genes (ISGs) implicated in regulation of protein translation. Poxvirus and coronavirus mutants that lacked 2′-O MTase activity similarly showed enhanced sensitivity to the antiviral actions of IFN and, specifically, IFIT proteins. Our results demonstrate that the 2′-O methylation of the 5′ cap of viral RNA functions to subvert innate host antiviral responses through escape of IFIT-mediated suppression, and suggest an evolutionary explanation for 2′-O methylation of cellular mRNA: to distinguish self from non-self RNA. Differential methylation of cytoplasmic RNA probably serves as an example for pattern recognition and restriction of propagation of foreign viral RNA in host cells.

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Figure 1: WNV-E218A is attenuated in wild-type mice and cells but is virulent in Ifnar1 −/− mice and cells.
Figure 2: 2′- O methylation of viral RNA alters the sensitivity of WNV to the antiviral effects of IFN.
Figure 3: WNV-E218A is more sensitive to the antiviral actions of Ifit genes.
Figure 4: Poxvirus and coronavirus mutants lacking 2′- O methylation are more sensitive to the antiviral effects of murine IFIT-2.


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This work was supported by National Institutes of Health grants U54 AI081680, U19 AI083019 and R01 AI074973 (to M.G. and M.S.D.), R01 AI56540 (to S.S.), U54 AI057160 (to R.M.B.) and U54 AI057158 (to P.Y.S.), R01 CA068782 (to G.C.S.) the Swiss National Science Foundation, 3100A0-118425/1, and the Novartis Foundation (to V.T. and R.Z.). We thank H. Virgin and B. Moss for reading the manuscript.

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S.D., K.J.S., R.M.B., T.C.P., M.G., P.-Y.S. and M.S.D. designed the experiments. S.D., K.J.S., J.S., J.L., T.-Y.L. and H.D. performed the experiments. S.D., K.J.S., J.S., R.M.B., M.G., P.-Y.S. and M.S.D. analysed the data. S.Y., V.F., G.C.S., W.B.K., R.Z. and V.T. provided key reagents and expertise. S.D., K.J.S., R.M.B, T.C.P., M.G., P.-Y.S. and M.S.D. wrote and edited the manuscript.

Correspondence to Michael S. Diamond.

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Daffis, S., Szretter, K., Schriewer, J. et al. 2′-O methylation of the viral mRNA cap evades host restriction by IFIT family members. Nature 468, 452–456 (2010) doi:10.1038/nature09489

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