Fig. 1: Visualization of actively translating E. coli polysomes.

a ¹⁴C-Val incorporation into nascent peptides over a 10-min period after incubation of 700 nM E. coli polysomes in the PURE system at 37 °C. Data (closed circles) represent mean values of triplicate experiments with error bars indicating the s. d. (n = 3). Data from duplicate experiments are shown individually (open cirles; t = 0 and t = 10 min). Data were fitted to Eqs. 1 and 2. b Translation cycle showing functional ribosome states isolated from ex vivo-derived polysomes reactivated in the PURE system. Shown are 30S (yellow), 50S (blue), initiation factors 1 (lavender), 2 (blue), and 3 (orange), translation factors (red), aminoacyl-tRNAs (A-tRNA, light violet), peptidyl-tRNAs (P-tRNA, green), and exit-tRNAs (E-tRNA, orange). For functional 70S state populations containing disome fractions, collided ribosomes are indicated as silhouettes. The EF-G bound translocation intermediate (Ti-POST: EF-G·GDP | P | E), not observed experimentally, is shown as transparent map simulated from PDB 7N2C²¹. All maps were filtered to 5 Å. c Distribution of 70S functional states of all imaged 70S (filled bars) and of 70S_L in disome complexes (dashed bars). Source data are provided as a Source Data file.