Abstract

Persisters are dormant phenotypic variants of bacterial cells that are tolerant to killing by antibiotics1. Persisters are associated with chronic infections and antibiotic treatment failure1,​2,​3. In Escherichia coli, toxin–antitoxin modules have been linked to persister formation4,​5,​6. The mechanism of persister formation in Gram-positive bacteria is unknown. Staphylococcus aureus is a major human pathogen, responsible for a variety of chronic and relapsing infections such as osteomyelitis, endocarditis and infections of implanted devices. Deleting toxin–antitoxin modules in S. aureus did not affect the level of persisters. Here, we show that S. aureus persisters are produced due to a stochastic entrance into the stationary phase accompanied by a drop in intracellular adenosine triphosphate. Cells expressing stationary-state markers are present throughout the growth phase, and increase in frequency with cell density. Cell sorting revealed that the expression of stationary markers is associated with a 100–1,000-fold increase in the likelihood of survival to antibiotic challenge. The adenosine triphosphate level of the cell is predictive of bactericidal antibiotic efficacy and explains bacterial tolerance to antibiotics.

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Acknowledgements

The authors thank C. Wolz for the gift of the HG001, HG001 rshsyn and triple mutant rshsyn, relP, relQ strains. The authors thank R. Lee and M. LaFleur for critical discussions. This work was supported by National Institutes of Health grant no. R01AI110578 to K.L. and by a Charles A. King fellowship to B.C.

Author information

Author notes

    • Brian P. Conlon
    •  & Sarah E. Rowe

    These authors contributed equally to this work.

Affiliations

  1. Antimicrobial Discovery Center, Department of Biology, Northeastern University, Boston, Massachusetts 02115, USA

    • Brian P. Conlon
    • , Sarah E. Rowe
    • , Autumn Brown Gandt
    • , Austin S. Nuxoll
    • , Eliza A. Zalis
    •  & Kim Lewis
  2. Synlogic, Cambridge, Massachusetts 02139, USA

    • Sarah E. Rowe
  3. Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth, Hanover, New Hampshire 03755, USA

    • Niles P. Donegan
    •  & Ambrose L. Cheung
  4. Biological Sciences Division, Pacific Northwest National Laboratory, Richland, Washington 99352, USA

    • Geremy Clair
    •  & Joshua N. Adkins

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Contributions

B.P.C. and S.E.R. designed the study, performed experiments, analysed results and wrote the paper. A.B.G., A.S.N. and E.A.Z. performed experiments. N.P.G. created the triple TA mutant strain. G.C. and J.N.A. designed the study and analysed results. A.L.C. designed the study. K.L. designed the study, analysed results and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Kim Lewis.

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https://doi.org/10.1038/nmicrobiol.2016.51

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