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The multiple antibiotic resistance regulator MarR is a copper sensor in Escherichia coli

Nature Chemical Biology volume 10, pages 21–28 (2014)Cite this article

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Abstract

The widely conserved multiple antibiotic resistance regulator (MarR) family of transcription factors modulates bacterial detoxification in response to diverse antibiotics, toxic chemicals or both. The natural inducer for Escherichia coli MarR, the prototypical transcription repressor within this family, remains unknown. Here we show that copper signaling potentiates MarR derepression in E. coli. Copper(II) oxidizes a cysteine residue (Cys80) on MarR to generate disulfide bonds between two MarR dimers, thereby inducing tetramer formation and the dissociation of MarR from its cognate promoter DNA. We further discovered that salicylate, a putative MarR inducer, and the clinically important bactericidal antibiotics norfloxacin and ampicillin all stimulate intracellular copper elevation, most likely through oxidative impairment of copper-dependent envelope proteins, including NADH dehydrogenase-2. This membrane-associated copper oxidation and liberation process derepresses MarR, causing increased bacterial antibiotic resistance. Our study reveals that this bacterial transcription regulator senses copper(II) as a natural signal to cope with stress caused by antibiotics or the environment.

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Figure 1: Copper(II) is a natural signal for MarR derepression.
Figure 2: The response mechanism of MarR to copper(II).
Figure 3: Crystal structure of copper(II)-oxidized MarR5CS(80C).
Figure 4: SAL- and antibiotic-triggered intracellular copper elevation.
Figure 5: Antibiotic-stimulated OHP production leads to copper liberation and MarR derepression.
Figure 6: A new linkage of antibiotic-triggered envelope stress and copper signaling with MarR-mediated bacterial antibiotic resistance.

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Acknowledgements

We thank J. Rosner (National Institute of Diabetes and Digestive and Kidney Diseases, US National Institutes of Health) for providing the strains of M2073 (WT E. coli) and M2076 (marRAB mutant strains) bearing the marR::lacZ reporter, J.J. Collins (Department of Biomedical Engineering, Boston University) for providing the pL(lexO)-GFP reporter, C.J. Chang (College of Chemistry, University of California–Berkeley) for providing the Cu(I)-specific fluorescent probe CS-1 and S.F. Reichard for editing. The E. coli WT (K12) strain (BW25113) and all of the single-gene deletion mutants were obtained from the National BioResource Project (National Institute of Genetics, Japan). We also thank the staff members of the Shanghai Synchrotron Radiation Facility and the Beijing Synchrotron Radiation Facility. This work was supported by research grants from the National Basic Research Foundation of China (2010CB912302 and 2012CB917301 to P.R.C.; 2011CB809103 to C.H.), the National Natural Science Foundation of China (21225206, 91013005 and 21001010 to P.R.C.), the US National Science Foundation (CHE-1213598 to C.H.) and the E-Institutes of Shanghai Municipal Education Commission (project number E09013 to C.H. and P.R.C.).

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Author notes

  1. Hubing Lou

    Present address: Current address: Cardiovascular Research Institute, University of California–San Francisco, San Francisco, California, USA.,

  2. Ziyang Hao and Hubing Lou: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemical Biology, Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, College of Chemistry and Molecular Engineering, Peking University, Beijing, China

    Ziyang Hao, Hubing Lou, Dianmu Zhang, Boxuan Simen Zhao, Xing Chen, Chuan He & Peng R Chen

  2. Peking-Tsinghua Center for Life Sciences, Beijing, China

    Rongfeng Zhu & Peng R Chen

  3. College of Biological Sciences, China Agricultural University, Beijing, China

    Jiuhe Zhu

  4. Department of Chemistry and Institute for Biophysical Dynamics, University of Chicago, Chicago, Illinois, USA

    Boxuan Simen Zhao & Chuan He

  5. International Curriculum Center, High School affiliated to Renmin University, Beijing, China

    Shizhe Zeng

  6. Department of Chemistry, University of California–Berkeley, Berkeley, California, USA

    Jefferson Chan

  7. Shanghai Universities E-Institute for Chemical Biology, Shanghai, China

    Chuan He & Peng R Chen

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Contributions

Z.H. performed the biochemical studies and participated in the structure studies. H.L. determined the crystal structures of copper(II)-oxidized MarR5CS(80C) and reduced MarRC80S. R.Z., J.Z. and X.C. helped with the biochemical and/or structure experiments. D.Z., B.S.Z. and S.Z. performed the experiments for OHP detection using OHSer. J.C. synthesized the Cu(I)-specific fluorescent probe CS-1. P.R.C. and C.H. conceived the study, designed experiments, interpreted data and wrote the manuscript with input from all of the authors.

Corresponding authors

Correspondence to Chuan He or Peng R Chen.

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Hao, Z., Lou, H., Zhu, R. et al. The multiple antibiotic resistance regulator MarR is a copper sensor in Escherichia coli. Nat Chem Biol 10, 21–28 (2014). https://doi.org/10.1038/nchembio.1380

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