The mechanism of error induction by the antibiotic viomycin provides insight into the fidelity mechanism of translation

• Main Authors: Mikael Holm, Chandra Sekhar Mandava, Måns Ehrenberg, Suparna Sanyal
• Format: Article
• Language: English
• Published: eLife Sciences Publications Ltd 2019-06-01
• Series: eLife
• Subjects: ribosome; protein synthesis; translation; antibiotic; viomycin
• Online Access: https://elifesciences.org/articles/46124

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.