FIG. 6.

Catalytic activities of EAV nsp11 mutants. (A) Purified N-terminally His₆-tagged EAV nsp11 proteins containing the indicated amino acid substitutions were electrophoresed on an SDS-polyacrylamide gel, which was subsequently stained with Coomassie brilliant blue. (B and C) Reaction mixtures containing 0.3 μM (B) or 1.5 μM (C) of mutant recombinant proteins and 2 μM of 5′-³²P-labeled RNA2 were incubated at 30°C. The reactions were stopped at the indicated time points, and the cleavage products were separated in a 20% polyacrylamide/7 M urea gel. For size markers, see the legend to Fig. 2. WT, wild type.