Nucleic Acids Research, 2002, Vol. 30, No. 9 1991-1996
© 2002 Oxford University Press
The p53-induced mouse zinc finger protein wig-1 binds double-stranded RNA with high affinity
1Karolinska Institute, Department of Oncology-Pathology, Cancer Center Karolinska (CCK), Karolinska Hospital, S-171 76 Stockholm, Sweden and 2Microbiology and Tumor Biology Center (MTC), Karolinska Institute, S-171 77 Stockholm, Sweden
The p53-induced mouse wig-1 gene encodes a Cys2His2-type zinc finger protein of unknown function. The zinc fingers in wig-1 are connected by long (5675) amino acid linkers. This distribution of zinc finger domains resembles that of the previously described double-stranded (ds)RNA-binding proteins dsRBP-ZFa and JAZ. Ectopically expressed FLAG-tagged mouse wig-1 protein localized to nuclei and in some cells to nucleoli, whereas GFP-tagged mouse wig-1 localized primarily to nucleoli. Electrophoretic mobility shift assay using a recombinant GSTwig-1 fusion protein showed that wig-1 preferentially binds dsRNA rather than single-stranded RNA or dsDNA. A set of deletion/truncation mutants of wig-1 was tested to determine the dsRNA-binding domain(s) or region(s) in wig-1 that is involved in the stabilization of wig-1dsRNA complexes in vitro. This revealed that the first zinc finger in wig-1 is essential for binding to dsRNA, whereas zinc fingers 2 and 3 are dispensable. wig-1 protein expressed in mammalian cells also showed a high affinity for dsRNA. wig-1 represents the first confirmed p53-induced gene that encodes a dsRNA-binding protein. This suggests that dsRNA binding plays a role in the p53-dependent stress response.
* To whom correspondence should be addressed at: Karolinska Institute, Department of Oncology-Pathology, Cancer Center Karolinska R8:04, S-171 76 Stockholm, Sweden. Tel: +46 8 51779342; Fax: +46 8 321047; Email: klas.wiman{at}mtc.ki.se Present address:Wang Qian, National Institute for Medical Research, Division of Virology, The Ridgeway, Mill Hill, London NW7 1AA, UK
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