TY - JOUR TI - The mechanism of error induction by the antibiotic viomycin provides insight into the fidelity mechanism of translation AU - Holm, Mikael AU - Mandava, Chandra Sekhar AU - Ehrenberg, Måns AU - Sanyal, Suparna A2 - Hinnebusch, Alan G A2 - Manley, James L VL - 8 PY - 2019 DA - 2019/06/07 SP - e46124 C1 - eLife 2019;8:e46124 DO - 10.7554/eLife.46124 UR - https://doi.org/10.7554/eLife.46124 AB - Applying pre-steady state kinetics to an Escherichia-coli-based reconstituted translation system, we have studied how the antibiotic viomycin affects the accuracy of genetic code reading. We find that viomycin binds to translating ribosomes associated with a ternary complex (TC) consisting of elongation factor Tu (EF-Tu), aminoacyl tRNA and GTP, and locks the otherwise dynamically flipping monitoring bases A1492 and A1493 into their active conformation. This effectively prevents dissociation of near- and non-cognate TCs from the ribosome, thereby enhancing errors in initial selection. Moreover, viomycin shuts down proofreading-based error correction. Our results imply a mechanism in which the accuracy of initial selection is achieved by larger backward rate constants toward TC dissociation rather than by a smaller rate constant for GTP hydrolysis for near- and non-cognate TCs. Additionally, our results demonstrate that translocation inhibition, rather than error induction, is the major cause of cell growth inhibition by viomycin. KW - ribosome KW - protein synthesis KW - translation KW - antibiotic KW - viomycin JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -