Figure 3.

Inhibition by SHFL of PRF and RT in vitro. RNAs derived from recoding reporter plasmids were translated in RRL in the presence of the indicated concentration of His-tagged SHFL or SHFL-M1. The addition of only water (W) or SHFL dilution buffer (DB) is also indicated. The products were resolved by 12% SDS-PAGE and visualized by autoradiography. ¹⁴C molecular size markers were also run on the gel (M). IFC represents an in-frame control translation of an mRNA in which the two reading frames are placed in-frame by the addition of a single nucleotide immediately downstream of the slippery sequence (see Table 1). Proteins generated by ribosomes that do not frameshift (stop) or that enter the −1 reading frame (PRF), or read through a stop codon (RT) are arrowed. In the EMCV and TMEV panels, the addition of the trans-activator protein 2A is shown by lane underlining.