期刊
VIROLOGY
卷 377, 期 1, 页码 1-6出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.virol.2008.04.026
关键词
flavivirus NS5; RNA cap methylation; flavivirus replication; West Nile virus; methyltransferase
类别
资金
- NIAID NIH HHS [U54-AI057158, AI061193, R21 AI065562-02, U01 AI061193, N01AI25490, U54 AI057158, U01 AI061193-05, AI25490] Funding Source: Medline
West Nile virus methyltransferase catalyzes N7 and 2'-O methylations of the viral RNA cap (GpppA-RNA -> m(7)GpppAm-RNA). The two methylation events are independent, as evidenced by efficient N7 methylation of GpppA-RNA -> m(7)GpppA-RNA and GpppAm-RNA -> m(7)GpppAm-RNA, and by the 2'-O methylation of GpppA-RNA -> GpppAm-RNA and m(7)GpppA-RNA -> m(7)GpppAm-RNA. However, the 2'-O methylation activity prefers substrate m(7)GpppA-RNA to GpppA-RNA, thereby determining the dominant methylation pathway as GpppARNA -> m(7)GpppA-RNA -> m(7)GpppAm-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 m(7)GpppAm-RNA, demonstrating that separate molecules of the enzyme can independently catalyze the two cap methylations in vitro. (c) 2008 Elsevier Inc. All rights reserved.
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