FIG. 6.

Specificity of the RNA-binding activity of rNSP5 measured by filter-binding assay. (A) Reaction mixtures containing 0.03 pmol of ³³P-labeled gene 8 mRNA, various amounts of unlabeled gene 8 dsRNA (47), and 2.7 pmol of either rNSP2 or rNSP5 were incubated for 30 min at room temperature. ³³P-labeled RNA bound to the protein was recovered by filtration through nitrocellulose membranes and quantified with a scintillation counter. The normalized values were plotted as a function of the molar ratio in mass of unlabeled to labeled nucleic acid in the reaction mixtures. (B and C) Same as in panel A except that the reaction mixtures contained 0.07 pmol of ³³P-labeled gene 8 mRNA, various amounts of unlabeled gene 8 mRNA, luciferase RNA (Promega), rotavirus genomic dsRNA or SacII-linearized pSP65g8(+) dsDNA, and 0.9 pmol of rNSP5.