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Compounds that select against the tetracycline-resistance efflux pump

Nature Chemical Biology volume 12, pages 902–904 (2016)Cite this article

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Abstract

We developed a competition-based screening strategy to identify compounds that invert the selective advantage of antibiotic resistance. Using our assay, we screened over 19,000 compounds for the ability to select against the TetA tetracycline-resistance efflux pump in Escherichia coli and identified two hits, β-thujaplicin and disulfiram. Treating a tetracycline-resistant population with β-thujaplicin selects for loss of the resistance gene, enabling an effective second-phase treatment with doxycycline.

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Figure 1: A high-throughput diffusion-based screen identifies compounds that select against tetracycline resistance.
Figure 2: β-Thujaplicin and disulfiram select for loss of tetracycline resistance.

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Acknowledgements

We thank D. Rudner, R. Mazitschek, and R. Moellering for helpful insights; J. Horn and J. Marchionna for custom screening plate manufacture and technical advice; D. Flood and S. Rudnicki for technical support in the primary screen; and J. Wang and J. Moore for technical support in the flow cytometry assay. All primary screening was performed at Harvard Medical School ICCB-L Screening Facility. This work was supported in part by National Institute of Allergy and Infectious Diseases grant U54 AI057159, US National Institutes of Health grants R01 GM081617 (to R.K.) and GM086258 (to J.C.), European Research Council FP7 ERC grant 281891 (to R.K.) and a National Science Foundation Graduate Fellowship (to L.K.S.).

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Authors and Affiliations

  1. Department of Systems Biology, Harvard Medical School, Boston, Massachusetts, USA

    Laura K Stone, Michael Baym, Tami D Lieberman, Remy Chait & Roy Kishony

  2. Institute of Science and Technology–Austria, Maria Gugging, 3400, Klosterneuburg, Austria

    Remy Chait

  3. Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts, USA

    Jon Clardy

  4. Department of Biology and Department of Computer Science, Technion–Israel Institute of Technology, Haifa, Israel

    Roy Kishony

Authors
  1. Laura K Stone
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  2. Michael Baym
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Contributions

L.K.S., J.C., and R.K. designed research; L.K.S. performed experiments and analyzed data; M.B. and R.C. built the imaging setup and M.B. developed the automation; L.K.S. and M.B. performed genomic sequencing; T.D.L. analyzed genomic sequencing data; R.C. contributed the initial plate and assay design; L.K.S. and R.K. wrote the manuscript.

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Correspondence to Roy Kishony.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Figures 1–14 and Supplementary Tables 1–6. (PDF 24325 kb)

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Stone, L., Baym, M., Lieberman, T. et al. Compounds that select against the tetracycline-resistance efflux pump. Nat Chem Biol 12, 902–904 (2016). https://doi.org/10.1038/nchembio.2176

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