Fig. 3.

Recombinant p33 promotes the self-cleavage activity of the satellite Tobacco ringspot virus ribozyme in vitro. (A) Schematic presentation of the RNA template carrying the ribozyme sequence before and after the self-cleavage. The satellite (−) ribozyme of TRSV (Buzayan et al., 1986, Fedor, 2000) forms an autocatalytic hairpin structure, which self-cleaves at the 3′ end TBSV RIV(+)RNA, releasing two RNA products as shown. (B) Representative denaturing gel of ³²P-labeled RNA cleavage products, which are derived from the template RNA synthesized by in vitro transcription with T7 RNA polymerase in the presence of 0, 1 and 2 μg of purified recombinant p33 are shown. Each experiment was repeated at least three times.