title: mRNA cap methylation influences pathogenesis of vesicular stomatitis virus in vivo.
abstract: One role of mRNA cap guanine-N-7 (G-N-7) methylation is to facilitate the efficient translation of mRNA. The role of mRNA cap ribose 2'-O methylation is enigmatic, although recent work has implicated this as a signature to avoid detection of RNA by the innate immune system (S. Daffis, K. J. Szretter, J. Schriewer, J. Q. Li, S. Youn, J. Errett, T. Y. Lin, S. Schneller, R. Zust, H. P. Dong, V. Thiel, G. C. Sen, V. Fensterl, W. B. Klimstra, T. C. Pierson, R. M. Buller, M. Gale, P. Y. Shi, M. S. Diamond, Nature 468:452-456, 2010, doi:10.1038/nature09489). Working with vesicular stomatitis virus (VSV), we previously showed that a panel of recombinant VSVs carrying mutations at a predicted methyltransferase catalytic site (rVSV-K1651A, -D1762A, and -E1833Q) or S-adenosylmethionine (SAM) binding site (rVSV-G1670A, -G1672A, and -G4A) were defective in cap methylation and were also attenuated for growth in cell culture. Here, we analyzed the virulence of these recombinants in mice. We found that rVSV-K1651A, -D1762A, and -E1833Q, which are defective in both G-N-7 and 2'-O methylation, were highly attenuated in mice. All three viruses elicited a high level of neutralizing antibody and provided full protection against challenge with the virulent VSV. In contrast, mice inoculated with rVSV-G1670A and -G1672A, which are defective only in G-N-7 methylation, were attenuated in vivo yet retained a low level of virulence. rVSV-G4A, which is completely defective in both G-N-7 and 2'-O methylation, also exhibited low virulence in mice despite the fact that productive viral replication was not detected in lung and brain. Taken together, our results suggest that abrogation of viral mRNA cap methylation can serve as an approach to attenuate VSV, and perhaps other nonsegmented negative-strand RNA viruses, for potential application as vaccines and viral vectors.
  
  
  OBJECTIVE
  Nonsegmented negative-sense (NNS) RNA viruses include a wide range of significant human, animal, and plant pathogens. For many of these viruses, there are no vaccines or antiviral drugs available. mRNA cap methylation is essential for mRNA stability and efficient translation. Our current understanding of mRNA modifications of NNS RNA viruses comes largely from studies of vesicular stomatitis virus (VSV). In this study, we showed that recombinant VSVs (rVSVs) defective in mRNA cap methylation were attenuated in vitro and in vivo. In addition, these methyltransferase (MTase)-defective rVSVs triggered high levels of antibody responses and provided complete protection against VSV infection. Thus, this study will not only contribute to our understanding of the role of mRNA cap MTase in viral pathogenesis but also facilitate the development of new live attenuated vaccines for VSV, and perhaps other NNS RNA viruses, by inhibiting viral mRNA cap methylation.
authors.affiliation: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Yuanmei
authors.initials: Y
authors.lastname: Ma
authors.forename: Yongwei
authors.initials: Y
authors.lastname: Wei
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhang
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hui
authors.initials: H
authors.lastname: Cai
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhu
authors.forename: Konstantin
authors.initials: K
authors.lastname: Shilo
authors.forename: Michael
authors.initials: M
authors.lastname: Oglesbee
authors.forename: Steven
authors.initials: S
authors.lastname: Krakowka
authors.forename: Sean P J
authors.initials: SP
authors.lastname: Whelan
authors.forename: Jianrong
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D001921:Brain / Q000473:pathology / Q000821:virology; D002460:Cell Line; D003673:Defective Viruses / Q000235:genetics / Q000378:metabolism; D005260:Female; D007928:Lethal Dose 50; D008168:Lung / Q000473:pathology / Q000821:virology; D008745:Methylation; D008780:Methyltransferases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010641:Phenotype; D012315:RNA Caps / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism*; D054243:Vesicular Stomatitis / Q000276:immunology / Q000473:pathology / Q000821:virology*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity; D019562:Viral Load; D014774:Virulence; D014779:Virus Replication
chemical_list: D000914:Antibodies, Viral; D012315:RNA Caps; D012333:RNA, Messenger; D012367:RNA, Viral; D008780:Methyltransferases