RNA helicase activity of Semliki Forest virus replicase protein NSP2

被引：134

作者：

de Cedrón, MG

Ehsani, N

Mikkola, ML

Garcia, JA

Kääriäinen, L

机构：

[1] Univ Helsinki, Inst Biotechnol, Bioctr Viikki, FIN-00014 Helsinki, Finland

[2] Univ Autonoma Madrid, Ctr Biol Mol, CSIC, E-28049 Madrid, Spain

来源：

FEBS LETTERS | 1999年 / 448卷 / 01期

基金：

芬兰科学院;

关键词：

alphavirus; RNA replication; helicase;

D O I：

10.1016/S0014-5793(99)00321-X

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Semliki Forest virus replicase protein nsP2 shares sequence homology with several putative NTPases and RNA helicases. NsP2 has RNA-dependent NTPase activity. Here we expressed polyhistidine-tagged nsP2 in Escherichia coli, purified it by metal-affinity chromatography, and used it in RNA helicase assays. RNA helicase CI of plum pox potyvirus was used as a positive control, Unwinding of alpha-P-32-labelled partially double-stranded RNA required nsP2, Mg2+ and NTPs, NsP2 with a mutation, K192N, in the NTP-binding sequence GVPGSGK(192)SA could not unwind dsRNA and had no NTPase activity. This is the first demonstration of RNA helicase activity within the large alphavirus superfamily, (C) 1999 Federation of European Biochemical Societies.

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页码：19 / 22

页数：4

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