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Oxidation of phosphorothioate DNA modifications leads to lethal genomic instability

Nature Chemical Biology volume 13, pages 888894 (2017) | Download Citation

Abstract

Genomic modification by sulfur in the form of phosphorothioate (PT) is widespread among prokaryotes, including human pathogens. Apart from its physiological functions, PT sulfur has redox and nucleophilic properties that suggest effects on bacterial fitness in stressful environments. Here we show that PTs are dynamic and labile DNA modifications that cause genomic instability during oxidative stress. In experiments involving isotopic labeling coupled with mass spectrometry, we observed sulfur replacement in PTs at a rate of 2% h−1 in unstressed Escherichia coli and Salmonella enterica. Whereas PT levels were unaffected by exposure to hydrogen peroxide (H2O2) or hypochlorous acid (HOCl), PT turnover increased to 3.8–10% h−1 after HOCl treatment and was unchanged by H2O2, consistent with the repair of HOCl-induced sulfur damage. PT-dependent sensitivity to HOCl extended to cytotoxicity and DNA strand breaks, which occurred at HOCl doses that were orders of magnitude lower than the corresponding doses of H2O2. The genotoxicity of HOCl in PT-containing bacteria suggests reduced fitness in competition with HOCl-producing organisms and during infections in humans.

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Acknowledgements

The authors gratefully acknowledge funding from the Deutsche Forschungsgemeinschaft (S.K.), the US National Science Foundation (CHE-1019990; P.C.D.), the US National Institute of Environmental Health Science (ES002109; P.C.D.), the US National Institute of Allergy and Infectious Diseases (AI112711; P.C.D.), the National Natural Science Foundation of China (31630002; D.Y.), and the Singapore-MIT Alliance for Research and Technology sponsored by the National Research Foundation of Singapore (P.C.D.).

Author information

Author notes

    • Brandon S Russell

    Present address: GlaxoSmithKline, Houston, Texas, USA.

    • Stefanie Kellner
    •  & Michael S DeMott

    These authors contributed equally to this work.

Affiliations

  1. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

    • Stefanie Kellner
    • , Michael S DeMott
    • , Ching Pin Cheng
    • , Brandon S Russell
    • , Bo Cao
    •  & Peter C Dedon
  2. State Key Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

    • Delin You
  3. Singapore-MIT Alliance for Research and Technology, Singapore.

    • Peter C Dedon

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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Peter C Dedon.

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DOI

https://doi.org/10.1038/nchembio.2407

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