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Journal of Virology, May 2008, p. 4295-4307, Vol. 82, No. 9
0022-538X/08/$08.00+0     doi:10.1128/JVI.02202-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

West Nile Virus Methyltransferase Catalyzes Two Methylations of the Viral RNA Cap through a Substrate-Repositioning Mechanism{triangledown}

Hongping Dong,1,2,{dagger} Suping Ren,3,{dagger} Bo Zhang,1 Yangsheng Zhou,3 Francesc Puig-Basagoiti,1 Hongmin Li,1,3 and Pei-Yong Shi1,3*

Wadsworth Center, New York State Department of Health, Albany, New York 12201,1 College of Life Science, Nanjing Normal University, People's Republic of China,2 Department of Biomedical Sciences, School of Public Health, State University of New York, Albany, New York 122013

Received 8 October 2007/ Accepted 20 February 2008

Flaviviruses encode a single methyltransferase domain that sequentially catalyzes two methylations of the viral RNA cap, GpppA-RNA->m7GpppA-RNA->m7GpppAm-RNA, by using S-adenosyl-L-methionine (SAM) as a methyl donor. Crystal structures of flavivirus methyltransferases exhibit distinct binding sites for SAM, GTP, and RNA molecules. Biochemical analysis of West Nile virus methyltransferase shows that the single SAM-binding site donates methyl groups to both N7 and 2'-O positions of the viral RNA cap, the GTP-binding pocket functions only during the 2'-O methylation, and two distinct sets of amino acids in the RNA-binding site are required for the N7 and 2'-O methylations. These results demonstrate that flavivirus methyltransferase catalyzes two cap methylations through a substrate-repositioning mechanism. In this mechanism, guanine N7 of substrate GpppA-RNA is first positioned to SAM to generate m7GpppA-RNA, after which the m7G moiety is repositioned to the GTP-binding pocket to register the 2'-OH of the adenosine with SAM, generating m7GpppAm-RNA. Because N7 cap methylation is essential for viral replication, inhibitors designed to block the pocket identified for the N7 cap methylation could be developed for flavivirus therapy.

* Corresponding author. Mailing address: 120 New Scotland Avenue, Albany, NY 12208. Phone: (518) 473-7487. Fax: (518) 473-1326. E-mail: ship{at}wadsworth.org

{triangledown} Published ahead of print on 27 February 2008.

{dagger} These authors made equal contributions.

Journal of Virology, May 2008, p. 4295-4307, Vol. 82, No. 9
0022-538X/08/$08.00+0     doi:10.1128/JVI.02202-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Zhang, B., Dong, H., Zhou, Y., Shi, P.-Y. (2008). Genetic Interactions among the West Nile Virus Methyltransferase, the RNA-Dependent RNA Polymerase, and the 5' Stem-Loop of Genomic RNA. J. Virol. 82: 7047-7058 [Abstract] [Full Text]  
  • Kroschewski, H., Lim, S. P., Butcher, R. E., Yap, T. L., Lescar, J., Wright, P. J., Vasudevan, S. G., Davidson, A. D. (2008). Mutagenesis of the Dengue Virus Type 2 NS5 Methyltransferase Domain. J. Biol. Chem. 283: 19410-19421 [Abstract] [Full Text]  


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