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Cryo-EM reveals multiple mechanisms of ribosome inhibition by doxycycline

Created on 11 Nov 2025

Authors

Stuart, W. S., Isupov, M. N., McLaren, M., Jenkins, C. H., Monier, A., Daum, B., Norville, I. H., Gold, V. A. M., Harmer, N. J.

Abstract

The widely prescribed antimicrobial doxycycline demonstrates a particularly low minimum inhibitory concentration against the zoonotic pathogen Coxiella burnetii. Doxycycline targets the bacterial ribosome, canonically blocking tRNA binding at the decoding center in the small subunit. Using cryo electron-microscopy, we analyzed doxycycline binding to C. burnetii and Escherichia coli ribosomes. Both structures reveal unexpected binding at the exit tunnel in the large subunit. In C. burnetii three doxycycline molecules stack to block the tunnel. In E. coli one doxycycline molecule triggers a major uncharacterized conformation of the ribosome that blocks tRNA binding. We identify a new ribosomal protein in the C. burnetii large subunit and characterize an additional prokaryotic ribosome hibernation promoting factor. These insights may aid the development of new ribosome inhibitor antibiotics.

Preprint server: bioRxiv
The authors list and abstract were imported from bioRxiv on 11 Nov 2025.

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