Extended Data Fig. 8: Processivity of HsDis3L2 on 7SL mimic and longer double-stranded RNA substrates.
a, Representative gels from pre-steady state nuclease titration assays with 1 nM 5′ P32 -radiolabeled RNA substrate and 50 nM HsDis3L2. The overall length of the species is indicated on the left of the panels. Each experiment was repeated independently with similar results for three concentrations of HsDis3L2. b, Representative gels from pulse-chase reactions of WT human Dis3L2 at 50 nM concentration and 1 nM radiolabeled RNA substrate. Cold chase was added to the reaction at the 3 min timepoint to a final concentration 5000 nM. Measurements were taken pre-chase at 3 min (pink dot), and post-chase at 4, 5, 7.5, and 10 min (blue gradient dots). Each experiment was repeated independently with similar results at two concentrations of HsDis3L2 with n = 2. c, d, e, RNA fold predictions and corresponding processivity values from pulse-chase experiments for 7SL-hairpinF-U16, 7SL-hairpinG-U16, and hairpinI-GCU14. Error bars mark the Standard Deviation of the Mean with n = 4.
