Abstract
The peptidoglycan cell wall is an essential structure for the growth of most bacteria. However, many are capable of switching into a wall-deficient L-form state in which they are resistant to antibiotics that target cell wall synthesis under osmoprotective conditions, including host environments. L-form cells may have an important role in chronic or recurrent infections. The cellular pathways involved in switching to and from the L-form state remain poorly understood. This work shows that the lack of a cell wall, or blocking its synthesis with β-lactam antibiotics, results in an increased flux through glycolysis. This leads to the production of reactive oxygen species from the respiratory chain, which prevents L-form growth. Compensating for the metabolic imbalance by slowing down glycolysis, activating gluconeogenesis or depleting oxygen enables L-form growth in Bacillus subtilis, Listeria monocytogenes and Staphylococcus aureus. These effects do not occur in Enterococcus faecium, which lacks the respiratory chain pathway. Our results collectively show that when cell wall synthesis is blocked under aerobic and glycolytic conditions, perturbation of cellular metabolism causes cell death. We provide a mechanistic framework for many anecdotal descriptions of the optimal conditions for L-form growth and non-lytic killing by β-lactam antibiotics.
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Data availability
The data that support the findings of this study are available from the corresponding author on request.
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Acknowledgements
We thank P. Cossart for the L. monocytogenes strains. All work in the Errington lab was funded by a European Research Council award (grant no. 670980). Work in the L.P.S.d.C. lab was supported by a Wellcome Trust New Investigator Award (104785/B/14/Z). The L.P.S.d.C. lab is also funded by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001060), the UK Medical Research Council (FC001060) and the Wellcome Trust (FC001060).
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Y.K. performed and analysed most of the experiments. R.M. and K.M. contributed strain constructions and preliminary genetic and microscopic experiments. A.S. and L.P.S.d.C. performed and analysed the metabolome and mass spectrometry experiments. All authors contributed to the experimental design and concepts. Y.K. and J.E. wrote the main text with contributions from all other authors.
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Supplementary Figs. 1–4, Legend for Supplementary Video 1, Supplementary Tables 1–3 and Supplementary References.
Supplementary Video 1
Contrasting effects of glucose and succinate on L-form death/growth phenotype (related to Fig. 3).
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Kawai, Y., Mercier, R., Mickiewicz, K. et al. Crucial role for central carbon metabolism in the bacterial L-form switch and killing by β-lactam antibiotics . Nat Microbiol 4, 1716–1726 (2019). https://doi.org/10.1038/s41564-019-0497-3
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DOI: https://doi.org/10.1038/s41564-019-0497-3
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