Fig 7. 2C^ATPase enhances hammerhead ribozyme activity.

(A) Schematic illustrations of the sequences and secondary structures of hammerhead ribozyme (purple) and its substrate RNA (red and blue). The ribozyme is a 58-nt unlabeled RNA synthesized by T7 RNA polymerase, and the 28-nt RNA substrate was synthesized with its 5′ end being HEX labeled. Under the correct structure of the hammerhead ribozyme, the cleavage of the substrate takes place between the 12^th and 13^th nucleotides from the 3′ end (indicated by arrow). (B) The RNA substrate was incubated with the ribozyme in the absence (lane 3) or presence (lanes 4–8) of increasing amounts (0.1–2 pmol) of MBP-2C^ATPase as indicated. The HEX-labeled 16-nt cleavage product (blue) was detected via denaturing gel electrophoresis and scanning. Non-ribozyme supplementation in the absence (lane 1) or presence (lane 2) of 2C^ATPase was used as a negative control. The non-cleaved substrate and cleaved product strands are indicated.