Structure and function of flavivirus NS5 methyltransferase

Yangsheng Zhou, Debashish Ray, Yiwei Zhao, Hongping Dong, Suping Ren, Zhong Li, Yi Guo, Kristen A. Bernard, Pei-Yong Shi, Hongmin Li

Research output: Contribution to journalArticle

220 Scopus citations

Abstract

The plus-strand RNA genome of flavivirus contains a 5′ terminal cap 1 structure (m7GpppAmG). The flaviviruses encode one methyltransferase, located at the N-terminal portion of the NS5 protein, to catalyze both guanine N-7 and ribose 2′-OH methylations during viral cap formation. Representative flavivirus methyltransferases from dengue, yellow fever, and West Nile virus (WNV) sequentially generate GpppA → m 7GpppA → m7GpppAm. The 2′-O methylation can be uncoupled from the N-7 methylation, since m7GpppA-RNA can be readily methylated to m7GpppAm-RNA. Despite exhibiting two distinct methylation activities, the crystal structure of WNV methyltransferase at 2.8 A resolution showed a single binding site for S-adenosyl-L-methionine (SAM), the methyl donor. Therefore, substrate GpppA-RNA should be repositioned to accept the N-7 and 2′-O methyl groups from SAM during the sequential reactions. Electrostatic analysis of the WNV methyltraasferase structure showed that, adjacent to the SAM-binding pocket, is a highly positively charged surface that could serve as an RNA binding site during cap methylations. Biochemical and mutagenesis analyses show that the N-7 and 2′-O cap methylations require distinct buffer conditions and different side chains within the K 61-D146-K182-E218 motif, suggesting that the two reactions use different mechanisms. In the context of complete virus, defects in both methylations are lethal to WNV; however, viruses defective solely in 27′-O methylation are attenuated and can protect mice from later wild-type WNV challenge. The results demonstrate that the N-7 methylation activity is essential for the WNV life cycle and, thus, methyltransferase represents a novel target for flavivirus therapy.

Original languageEnglish (US)
Pages (from-to)3891-3903
Number of pages13
JournalJournal of Virology
Volume81
Issue number8
DOIs
StatePublished - Apr 2007
Externally publishedYes

    Fingerprint

ASJC Scopus subject areas

  • Immunology

Cite this

Zhou, Y., Ray, D., Zhao, Y., Dong, H., Ren, S., Li, Z., Guo, Y., Bernard, K. A., Shi, P-Y., & Li, H. (2007). Structure and function of flavivirus NS5 methyltransferase. Journal of Virology, 81(8), 3891-3903. https://doi.org/10.1128/JVI.02704-06