Figure 3

Cleavage kinetics using ribo-modified substrate. (A) Time course (0.25-60 minutes) of the cleavage reaction of purified wild type (lanes 1–8), and Glu710Gly mutant (lanes 9–16) with the CL14-U/CP25 substrate (shown at the top of the figure). In lane 17 the protein has not been added. CL1 represents the DNA strand cleaved by the enzymes at the preferred cleavage site, indicated by an arrow at the top of the figure. (B) Percentage of cleaved substrate, normalized to the maximum value of the wild type, plotted against time for the reaction with wild type (black diamond’s) and with Glu710Gly mutant (grey squares). Data shown are means ± SD from 3 independent experiments.