Figure 1: Cryo-EM structure of the ErmBL-stalled ribosome complex. | Nature Communications

Figure 1: Cryo-EM structure of the ErmBL-stalled ribosome complex.

From: Molecular basis for erythromycin-dependent ribosome stalling during translation of the ErmBL leader peptide

Figure 1

(a) Schematic for ermBL-dependent regulation of ermB translation in the presence of erythromycin (ERY). (b) In vitro translation of ErmBL using either 14C-Asp or 14C-Lys in the absence (−) or presence (E) of 50 μM ERY. In the control samples (B), where translation was carried out in the absence of ERY, but in presence of borrelidin (a Thr-tRNA synthetase inhibitor), translation arrest occurred after an 11-amino-acid nascent chain was polymerized (such that codon 12 is in the A-site, which is one codon further compared with the natural stall site), thereby providing a mobility marker and confirming14C-Lys incorporation in the absence of ERY. (c) The bicistronic 2 × ermBL mRNA was translated in vitro in the presence of 10 μM ERY to generate ErmBL-SRC disomes. (d) Complementary DNA oligo and RNase H cleavage converts disomes to monosomes, as shown by sucrose density centrifugation and negative stain EM. (e) Surface and cross-section of the ErmBL-SRC, containing 30S (yellow), 50S (coloured according to local resolution), A-tRNA (orange), P-tRNA (green) and ERY (red). Inset shows electron density for ERY (grey mesh) with fitted crystal structure (PDB3OFR)11.

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