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Persister formation in Staphylococcus aureus is associated with ATP depletion

Nature Microbiology volume 1, Article number: 16051 (2016) Cite this article

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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 failure1–3. In Escherichia coli, toxin–antitoxin modules have been linked to persister formation4–6. 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.

Antibiotic resistance is a major human health problem7. However, most pathogens that cause hard-to-treat chronic infections are not drug-resistant1,3,8. There is mounting evidence that drug-tolerant persister cells contribute to this phenomenon2,9–12. Persister cells are phenotypic variants that survive lethal doses of antibiotics and are genetically identical to their drug-susceptible kin. The mechanism of persister formation has been extensively studied in the closely related Gram-negative organisms Escherichia coli and Salmonella Typhimurium1,13,14.

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Figure 1: Toxin–antitoxin modules and stringent response do not control persister formation in S. aureus.
Figure 2: Activation of stationary markers is heterogeneous.
Figure 3: Persister sorting using stationary markers Pcap5A and ParcA.
Figure 4: Reduction in ATP induces persister formation and expression of stationary-phase markers.

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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.

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Author notes

  1. Brian P. Conlon and Sarah E. Rowe: These authors contributed equally to this work.

Authors and Affiliations

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

    Brian P. Conlon, Sarah E. Rowe, Autumn Brown Gandt, Austin S. Nuxoll, Eliza A. Zalis & Kim Lewis

  2. Synlogic, Cambridge, 02139, Massachusetts, USA

    Sarah E. Rowe

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

    Niles P. Donegan & Ambrose L. Cheung

  4. Biological Sciences Division, Pacific Northwest National Laboratory, Richland, 99352, Washington, 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.

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Correspondence to Kim Lewis.

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The authors declare no competing financial interests.

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Conlon, B., Rowe, S., Gandt, A. et al. Persister formation in Staphylococcus aureus is associated with ATP depletion. Nat Microbiol 1, 16051 (2016). https://doi.org/10.1038/nmicrobiol.2016.51

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