Separate molecules of West Nile virus methyltransferase can independently catalyze the N7 and 2′-O methylations of viral RNA cap

Hongping Dong, Suping Ren, Hongmin Li, Pei-Yong Shi

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

West Nile virus methyltransferase catalyzes N7 and 2′-O methylations of the viral RNA cap (GpppA-RNA → m7GpppAm-RNA). The two methylation events are independent, as evidenced by efficient N7 methylation of GpppA-RNA → m7GpppA-RNA and GpppAm-RNA → m7GpppAm-RNA, and by the 2′-O methylation of GpppA-RNA → GpppAm-RNA and m7GpppA-RNA → m7GpppAm-RNA. However, the 2′-O methylation activity prefers substrate m7GpppA-RNA to GpppA-RNA, thereby determining the dominant methylation pathway as GpppA-RNA → m7GpppA-RNA → m7GpppAm-RNA. Mutant enzymes with different methylation defects can trans complement one another in vitro. Furthermore, sequential treatment of GpppA-RNA with distinct methyltransferase mutants generates fully methylated m7GpppAm-RNA, demonstrating that separate molecules of the enzyme can independently catalyze the two cap methylations in vitro.

Original languageEnglish (US)
Pages (from-to)1-6
Number of pages6
JournalVirology
Volume377
Issue number1
DOIs
StatePublished - Jul 20 2008
Externally publishedYes

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RNA Caps
West Nile virus
Viral RNA
Methyltransferases
Methylation
RNA

Keywords

  • Flavivirus NS5
  • Flavivirus replication
  • Methyltransferase
  • RNA cap methylation
  • West Nile virus

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Separate molecules of West Nile virus methyltransferase can independently catalyze the N7 and 2′-O methylations of viral RNA cap. / Dong, Hongping; Ren, Suping; Li, Hongmin; Shi, Pei-Yong.

In: Virology, Vol. 377, No. 1, 20.07.2008, p. 1-6.

Research output: Contribution to journalArticle

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abstract = "West Nile virus methyltransferase catalyzes N7 and 2′-O methylations of the viral RNA cap (GpppA-RNA → m7GpppAm-RNA). The two methylation events are independent, as evidenced by efficient N7 methylation of GpppA-RNA → m7GpppA-RNA and GpppAm-RNA → m7GpppAm-RNA, and by the 2′-O methylation of GpppA-RNA → GpppAm-RNA and m7GpppA-RNA → m7GpppAm-RNA. However, the 2′-O methylation activity prefers substrate m7GpppA-RNA to GpppA-RNA, thereby determining the dominant methylation pathway as GpppA-RNA → m7GpppA-RNA → m7GpppAm-RNA. Mutant enzymes with different methylation defects can trans complement one another in vitro. Furthermore, sequential treatment of GpppA-RNA with distinct methyltransferase mutants generates fully methylated m7GpppAm-RNA, demonstrating that separate molecules of the enzyme can independently catalyze the two cap methylations in vitro.",
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