FIGURE 6.

The CTFV stem–loop stimulatory RNA does not induce ribosomal pausing. Messenger RNAs derived from AvaII-cut pPS0-CTFV-106-CGA were translated in RRL for 3 min at 26°C prior to addition of edeine to 5 μM. Aliquots were removed at various times post-edeine addition, translation stopped, and products resolved on a 10% SDS–polyacrylamide gel. As a positive control, the experiment was also carried out with an mRNA derived from AvaII-cut pPS1a, which contains the frameshift-promoting pseudoknot of the coronavirus IBV at the equivalent position in the mRNA as the CTFV stimulatory RNA in pPS0-CTFV-106-CGA. The expected size of the pausing product (Pex; shown by an asterisk) was marked by translating RNA produced from XhoI-cleaved pPS0 (C2). Control C1 represents the translation product produced from the mRNAs in the absence of added edeine.