Figure 3.
Kinetic profiles of 2′,3′-cyclic phosphate and 3′-phosphate ligation. Reaction mixtures containing 50 mM Tris-HCl (pH 8.0), 2 mM MnCl2, 0.1 mM GTP, 1 µM RtcB and 0.1 µM HORNA > p (open circle) or HORNAp (closed circle) substrates were incubated at 37°C. The reactions were quenched with EDTA at the times specified. The RNAs were digested with RNase T1 and analyzed by Urea–PAGE. The extent of ligation is plotted as a function of time. Each datum is the average of three independent experiments ± SEM.