FIG 7.
Mutational effects on HORNA>p ligation. (A) Reaction mixtures (10 μl) containing 50 mM Tris-HCl (pH 8.0), 2 mM MnCl2, 100 μM GTP, 0.1 μM 20-mer HORNA>p (depicted at the bottom, with the radiolabeled phosphate denoted by ●), and either 1 μM RtcB as specified or no RtcB (lane −) were incubated at 37°C for 5 min. The mixtures were digested with RNase T1 and then analyzed by urea-PAGE. An autoradiograph of the gel is shown. The identities of the radiolabeled RNase T1 fragments are indicated at the left and right. (B to D) The extents of RNA ligation (HOCUUpCpUGp) (B) and accumulation of RNAp (HOCUUpCp) (C) and RNAppG (HOCUUpCppG) (D), expressed as a percentage of the total radiolabeled RNA, are plotted in bar graph format. Each datum is the average of results from three experiments ± the standard error of the mean.