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Initiation factors rectify mRNA positioning in 70S complexes. Ribosomes (2 µM, CUGG; 1 µM, AUGG) were incubated with fMet-tRNAfMet2M1 or fMet-tRNAfMet2 (2 µM, as indicated) 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.
