Biochemical and genetic characterization of dengue virus methyltransferase

Hongping Dong, David C. Chang, Xuping Xie, Ying Xiu Toh, Ka Yan Chung, Gang Zou, Julien Lescar, Siew Pheng Lim, Pei-Yong Shi

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

We report that dengue virus (DENV) methyltransferase sequentially methylates the guanine N-7 and ribose 2'-O positions of viral RNA cap (GpppA→m7GpppA→m7GpppAm). The order of two methylations is determined by the preference of 2'-O methylation for substrate m7GpppA-RNA to GpppA-RNA, and the 2'-O methylation is not absolutely dependent on the prior N-7 methylation. A mutation that completely abolished the 2'-O methylation attenuated DENV replication in cell culture, whereas another mutation that abolished both methylations was lethal for viral replication, suggesting that N-7 methylation is more important than 2'-O methylation in viral replication. The latter mutant with lethal replication could be rescued by trans complementation using a wild-type DENV replicon. Furthermore, we found that chimeric DENVs containing the West Nile virus methyltransferase, polymerase, or full-length NS5 were nonreplicative, but the replication defect could also be rescued through trans complementation using the wild-type DENV replicon.

Original languageEnglish (US)
Pages (from-to)568-578
Number of pages11
JournalVirology
Volume405
Issue number2
DOIs
StatePublished - Sep 30 2010
Externally publishedYes

Fingerprint

Dengue Virus
Methyltransferases
Methylation
Molecular Biology
Replicon
RNA Caps
RNA
West Nile virus
Mutation
Ribose
Viral RNA
Guanine
Virus Replication
Cell Culture Techniques

Keywords

  • Antiviral target
  • Dengue virus
  • Flavivirus replication
  • Methyltransferase
  • RNA cap methylation

ASJC Scopus subject areas

  • Virology

Cite this

Dong, H., Chang, D. C., Xie, X., Toh, Y. X., Chung, K. Y., Zou, G., ... Shi, P-Y. (2010). Biochemical and genetic characterization of dengue virus methyltransferase. Virology, 405(2), 568-578. https://doi.org/10.1016/j.virol.2010.06.039

Biochemical and genetic characterization of dengue virus methyltransferase. / Dong, Hongping; Chang, David C.; Xie, Xuping; Toh, Ying Xiu; Chung, Ka Yan; Zou, Gang; Lescar, Julien; Lim, Siew Pheng; Shi, Pei-Yong.

In: Virology, Vol. 405, No. 2, 30.09.2010, p. 568-578.

Research output: Contribution to journalArticle

Dong, H, Chang, DC, Xie, X, Toh, YX, Chung, KY, Zou, G, Lescar, J, Lim, SP & Shi, P-Y 2010, 'Biochemical and genetic characterization of dengue virus methyltransferase', Virology, vol. 405, no. 2, pp. 568-578. https://doi.org/10.1016/j.virol.2010.06.039
Dong H, Chang DC, Xie X, Toh YX, Chung KY, Zou G et al. Biochemical and genetic characterization of dengue virus methyltransferase. Virology. 2010 Sep 30;405(2):568-578. https://doi.org/10.1016/j.virol.2010.06.039
Dong, Hongping ; Chang, David C. ; Xie, Xuping ; Toh, Ying Xiu ; Chung, Ka Yan ; Zou, Gang ; Lescar, Julien ; Lim, Siew Pheng ; Shi, Pei-Yong. / Biochemical and genetic characterization of dengue virus methyltransferase. In: Virology. 2010 ; Vol. 405, No. 2. pp. 568-578.
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