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Ubiquinone accumulation improves osmotic-stress tolerance in Escherichia coli

Nature Chemical Biology volume 10, pages 266–272 (2014)Cite this article

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Abstract

Bacteria are thought to cope with fluctuating environmental solute concentrations primarily by adjusting the osmolality of their cytoplasm. To obtain insights into the underlying metabolic adaptations, we analyzed the global metabolic response of Escherichia coli to sustained hyperosmotic stress using nontargeted mass spectrometry. We observed that 52% of 1,071 detected metabolites, including known osmoprotectants, changed abundance with increasing salt challenge. Unexpectedly, unsupervised data analysis showed a substantial increase of most intermediates in the ubiquinone-8 (Q8) biosynthesis pathway and a 110-fold accumulation of Q8 itself, as confirmed by quantitative lipidomics. We then demonstrated that Q8 is necessary for acute and sustained osmotic-stress tolerance using Q8 mutants and tolerance rescue through feeding nonrespiratory Q8 analogs. Finally, in vitro experiments with artificial liposomes showed that mechanical membrane stabilization is a principal mechanism of Q8-mediated osmoprotection. Thus, we find that besides regulating intracellular osmolality, E. coli enhances its cytoplasmic membrane stability to withstand osmotic stress.

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Figure 1: Analysis of the salt-induced metabolic changes in E. coli by mass spectrometry.
Figure 2: The global metabolic response of E. coli to hyperosmotic stress.
Figure 3: The ubiquinone pool and its precursors increase upon osmotic stress.
Figure 4: Ubiquinone is required for osmotic-stress tolerance in vivo.
Figure 5: Ubiquinone enhances the stability of artificial liposomes.

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Acknowledgements

We thank N. Zamboni for advice and assistance with MS/MS analyses, A. Kühne for assistance with microscopy and K. Kochanowski and T. Fuhrer for helpful discussions. Funding was provided by the MetaNetX project of the Swiss Initiative for Systems Biology (SystemsX.ch; http://metanetx.org/) evaluated by the Swiss National Science Foundation, and the Swiss Federal Government through the Federal Office of Education and Science.

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Authors and Affiliations

  1. Institute of Molecular Systems Biology, Swiss Federal Institute of Technology (ETH) Zürich, Zürich, Switzerland

    Daniel C Sévin & Uwe Sauer

  2. PhD Program on Systems Biology, Life Science Zürich, Zürich, Switzerland

    Daniel C Sévin

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  1. Daniel C Sévin
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D.C.S. performed the experiments and analyzed the data. D.C.S. and U.S. designed the research and wrote the paper.

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Correspondence to Uwe Sauer.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Results and Supplementary Figures 1–7. (PDF 2894 kb)

Supplementary Data Sets 1–3

Supplementary Data Set 1 contains the intensity values of annotated ions in the metabolomics data set, normalized and log2-transformed to the 50 mM condition and filtered for ions showing at least a 2-fold change in the 500 mM condition. Supplementary Data Set 2 lists the metabolite annotation of each ion in the metabolomics experiments (Supplementary Data Set 1), with the electrospray modification indicated in square brackets. All metabolites differ less than 0.001 Da from the theoretical mass. The KEGG eco metabolite database was used as annotation reference. Supplementary Data Set 3 contains the intensity values of ions detected in the the lipidomics experiment, shown as fold-change values of the 450 vs. 50 mM NaCl experiments and filtered for ions showing at least a 2-fold change. P-values were calculated by two-sided t-tests assuming unequal variance. Additionally, lipid annotations of the ions are listed with electrospray modifications given in square brackets. Lipid ions differ less than 0.001 Da from the theoretical mass in the KEGG eco metabolite database. (XLSX 65 kb)

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Sévin, D., Sauer, U. Ubiquinone accumulation improves osmotic-stress tolerance in Escherichia coli. Nat Chem Biol 10, 266–272 (2014). https://doi.org/10.1038/nchembio.1437

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