Figure 6.
Role of protons in tRNA translocation. (A) RIC-liposomes (25 μg/ml) were washed and incubated in buffers of indicated pH in the absence or presence of 4 mM ATP, with 1 nM of BU-labeled tRNA. The vesicles containing the bound RNAs were then transferred to 37°C for 10 min and then UV crosslinked. (B) BU-labeled tRNATyr was bound to RIC-liposomes which had been pre-incubated with 50 μM of either m-chlorocarbonylcyanide phenylhydrazone (CCCP) or oligomycin, and incubated at 37°C in the presence of 4 mM ATP for 2 or 10 min. (C) Effect of valinomycin (50 μM) plus 50 mM KCl, or nigericin (50 μM) on translocation. Reaction conditions were as in (B).