Figure 2.
Effect of Archease on the single-turnover rate of RNA ligation by RtcB. (A) RNA ligation reactions with RtcB alone or with the inclusion of 100 nM Archease. Reaction mixtures contained 50 mM Bis–Tris buffer (pH 7.0), NaCl (300 mM), MnCl2 (0.25 mM), GTP (0.10 mM), P. horikoshii RtcB (5 μM), 5′ RNA fragment (1.0 μM) and 3′ RNA fragment (1.0 μM). Reaction mixtures were incubated at 70°C, and aliquots were removed at the indicated times and quenched with an equal volume of RNA gel-loading buffer. (B) Plots of ligation product formation over time fitted to a single-exponential equation. Values are the mean ± SE for two separate experiments.