Figure 7.

Initiation factors rectify mRNA positioning in various 30S complexes. 30S subunits (2 µM, CUGG; 1 µM, AUGG) were incubated with fMet-tRNA^fMet2M1 (A-B, 2 µM), tRNA^fMet2M1 (C, 2 µM), or fMet-tRNA^fMet2 (D-E, 2 µM) in the presence of mRNA (CUGG, 1 µM; AUGG, 0.1 µM), GTP (100 µM), and in the absence or presence of initiation factors (3 µM each, as indicated) at 37°C for 5 min, and complexes were analyzed by toeprinting. Histograms show the distribution of toeprint signal versus toeprint position for complexes containing mutant (B) or control (E) tRNA. Data represent the mean ± SEM from ≥ 3 independent experiments. The dotted red line benchmarks position +16.