Fig 8. The CTD of 2C^ATPase is required for its RNA chaperone activity.

(A) The standard RNA helix (0.1 pmol) was reacted with MBP-fusion 2C^ATPase Wt (lane 3), GK134AA mutant (lane 4), or a CTD fragment (lane 5). Native (lane 1) or boiled reaction mixture (lane 2) was used as a negative or positive control, respectively. (B) The ATPase activity of the indicated proteins was measured as nanomoles of released inorganic phosphate as indicated. MBP alone was used as the negative control. Error bars represent SD values from three separate experiments. (C) and (D) The standard RNA helix (0.1 pmol) was reacted with MBP-2C^ATPase Wt (lane 3), GK134AA mutant (lane 5), or ΔCTD (lane 4) in the absence (C) or presence (D) of 5 mM ATP as indicated. (E) The unwinding activities of Wt or indicated mutant 2C^ATPase in the absence or presence of 5 mM ATP were plotted as the percentage of the released RNA from the total RNA helix substrate. Error bars represent SD values from three separate experiments.