Journal of General Virology, Vol 80, 1119-1126, Copyright © 1999 by Society for General Microbiology

ARTICLES

Identification of the RNA-binding sites of the triple gene block protein 1 of bamboo mosaic potexvirus

CH Wung, YH Hsu, DY Liou, WC Huang, NS Lin and BY Chang
Agricultural Biotechnology Laboratories, National Chung-Hsing University, Taichung, Taiwan 402, Republic of China

The triple gene block protein 1 (TGBp1) encoded by open reading frame 2 of bamboo mosaic potexvirus (BaMV) was overexpressed in Escherichia coli and purified in order to test its RNA-binding activity. UV crosslinking assays revealed that the RNA-binding activity was present mainly in the soluble fraction of the refolded TGBp1. The binding activity was nonspecific and salt concentration-dependent: activity was present at 0--50 mM NaCl but was almost abolished at 200 mM. The RNA-binding domain was located by deletion mutagenesis to the N-terminal 3--24 amino acids of TGBp1. Sequence alignment analysis of the N-terminal 25 amino acids of the TGBp1 homologues of potexviruses identified three arginine residues. Arg-to-Ala substitution at any one of the three arginines eliminated most of the RNA-binding activity, indicating that they were all critical to the RNA-binding activity of the TGBp1 of BaMV.

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