Fig. 2: AzAt Photoaffinity Labeling.

a AzAt photoincorporation into Stop-POST5, the RNA fraction of Stop-POST5, 80S.IRES, and the RNA fraction of 80S.IRES. All of the labeling stoichiometries are normalized to that of Stop-POST5 at 600 µM AzAt, which was equal to 1.2/Stop-POST5. b AzAt photoincorporation into eRF1 both alone and complexed with eRF3.GDPNP. The values are normalized to the saturation labeling of isolated eRF1. c Inhibition of AzAt (250 µM) photoincorporation by the addition of either ataluren (1000 µM) or GJ072 (150 µM). d The mutation rate fold change for PAL vs. PRE samples for Stop-POST5 (red) and 80S.IRES complexes (blue) for the 22 sites most pertinent for ataluren function, and the asterisks indicates sites of particular interest (see text). e Saturation curves for photoincorporation into 18S-A1195, as measured by photoincorporation into Fragment I, vs. the sum of the photoincorporations into 26S A3093, A2669, and A2672, as measured by photoincorporation into Fragments II and III. f Location of A1195 within the 40S subunit containing bound eRF1. g Locations of A2669, A2672, and A3093 within the 60S subunit containing bound eRF1. All of the error bars in this Figure represent average deviations for n = 2 independent determinations, with the exception of the eRF1 labeling within the ternary complex in b, for which n=3. Source data for (a–e) are provided as a Source Data file.