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Identification and characterization of the RNA helicase activity of Japanese encephalitis virus NS3 protein. 2000
A Utama, and
H Shimizu, and
S Morikawa, and
F Hasebe, and
K Morita, and
A Igarashi, and
M Hatsu, and
K Takamizawa, and
T Miyamura
Department of Bioprocessing, Faculty of Agriculture, Gifu University, 1-1 Yanagido, Gifu, Japan.
The NS3 protein of Japanese encephalitis virus (JEV) contains motifs typical of RNA helicase/NTPase but no RNA helicase activity has been reported for this protein. To identify and characterize the RNA helicase activity of JEV NS3, a truncated form of the protein with a His-tag was expressed in Escherichia coli and purified. The purified JEV NS3 protein showed an RNA helicase activity, which was dependent on divalent cations and ATP. An Asp-285-to-Ala substitution in motif II of the JEV NS3 protein abolished the ATPase and RNA helicase activities. These results indicate that the C-terminal 457 residues are sufficient to exhibit the RNA helicase activity of JEV NS3.
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