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Identification and characterization of the RNA helicase activity of Japanese encephalitis virus NS3 protein
Author(s) -
Utama Andi,
Shimizu Hiroyuki,
Morikawa Shigeru,
Hasebe Futoshi,
Morita Kouichi,
Igarashi Akira,
Hatsu Masahiro,
Takamizawa Kazuhiro,
Miyamura Tatsuo
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)01705-6
Subject(s) - helicase , rna helicase a , ns3 , rna , degradosome , rna dependent rna polymerase , virology , japanese encephalitis , biology , microbiology and biotechnology , virus , chemistry , biochemistry , encephalitis , gene , hepatitis c virus
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.