West Nile virus methyltransferase catalyzes two methylations of the viral RNA cap through a substrate-repositioning mechanism

Hongping Dong, Suping Ren, Bo Zhang, Yangsheng Zhou, Francesc Puig-Basagoiti, Hongmin Li, Pei-Yong Shi

Research output: Contribution to journalArticle

74 Citations (Scopus)

Abstract

Flaviviruses encode a single methyltransferase domain that sequentially catalyzes two methylations of the viral RNA cap, GpppA-RNA→m 7GpppA-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 m 7GpppA-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.

Original languageEnglish (US)
Pages (from-to)4295-4307
Number of pages13
JournalJournal of Virology
Volume82
Issue number9
DOIs
StatePublished - May 2008
Externally publishedYes

Fingerprint

RNA Caps
West Nile virus
Viral RNA
methyltransferases
Methyltransferases
S-Adenosylmethionine
methylation
Methylation
Flavivirus
S-adenosylmethionine
RNA
Guanosine Triphosphate
Binding Sites
binding sites
Guanine
Flaviviridae
Adenosine
guanine
adenosine
virus replication

ASJC Scopus subject areas

  • Immunology

Cite this

West Nile virus methyltransferase catalyzes two methylations of the viral RNA cap through a substrate-repositioning mechanism. / Dong, Hongping; Ren, Suping; Zhang, Bo; Zhou, Yangsheng; Puig-Basagoiti, Francesc; Li, Hongmin; Shi, Pei-Yong.

In: Journal of Virology, Vol. 82, No. 9, 05.2008, p. 4295-4307.

Research output: Contribution to journalArticle

Dong, Hongping ; Ren, Suping ; Zhang, Bo ; Zhou, Yangsheng ; Puig-Basagoiti, Francesc ; Li, Hongmin ; Shi, Pei-Yong. / West Nile virus methyltransferase catalyzes two methylations of the viral RNA cap through a substrate-repositioning mechanism. In: Journal of Virology. 2008 ; Vol. 82, No. 9. pp. 4295-4307.
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