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Nitric oxide upregulates expression of DNA-PKcs to protect cells from DNA-damaging anti-tumour agents

Nature Cell Biology volume 2, pages 339–345 (2000)Cite this article

Abstract

Nitric-oxide synthase (NOS) activity has been detected in many human tumours, although its function is unclear. Here we show that exposure of cells to nitric oxide (NO) results in a 4–5-fold increase in expression of the DNA-dependent protein-kinase catalytic subunit (DNA-PKcs), one of the key enzymes involved in repairing double-stranded DNA breaks. This NO-mediated increase in enzymatically active DNA-PK not only protects cells from the toxic effects of NO, but also provides crossprotection against clinically important DNA-damaging agents, such as X-ray radiation, adriamycin, bleomycin and cisplatin. The NO-mediated increase in DNA-PKcs described here demonstrates the presence of a new and highly effective NO-mediated mechanism for DNA repair.

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Figure 1: Generation of NO by EcR293 clone-11 cells.
Figure 2: Expression of DNA-PKcs mRNA.
Figure 3: Expression of DNA-PKcs.
Figure 4: DNA‐PK pulldown peptide assay.
Figure 5: NO-dependent increase in DNA-PKcs promoter activity.
Figure 6: NO-mediated increase in Sp1 binding.
Figure 7: NO-generating cells are protected from DNA-damaging agents.
Figure 8: Cytokine induction of NO or exogenous administration of an NO donor protects cells from DNA damaging agents.

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Acknowledgements

We thank C. Jenkins and S. Jackson for discussions, C. W. Anderson (Brookhaven National Laboratory, New York) for DNA-PKcs promoter–reporter plasmids, R. Tjian (Howard Hughes Medical Institute, Berkeley, Illinois) for pPacSp1 and pPacO plasmids and M. J. Allalunis-Turner for cell lines M059J and M059K. We also thank D. Sheer for help in the use of the X-ray facility at the ICRF, London. This work was supported by a grant from the Medical Research Council.

Correspondence and requests for materials should be addressed to I.G.C.

Supplementary information is available on Nature Cell Biology’s website (http://www.nature.com/ncb) or as paper copy from the London editorial office of Nature Cell Biology.

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Author notes

  1. Graeme C. M. Smith

    Present address: KuDOS Pharmaceuticals Ltd, Cambridge, CB4 4WG, UK

Authors and Affiliations

  1. The Wolfson Institute for Biomedical Research, The Cruciform Building, University College London, Gower Street, London, WC1E 6BT, UK

    Weiming Xu, Lizhi Liu & lan G. Charles

  2. Wellcome Trust/CRC Institute and Department of Zoology, Cambridge University, Tennis Court Road, Cambridge, CB2 1QR, UK

    Graeme C. M. Smith

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Correspondence to lan G. Charles.

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Xu, W., Liu, L., Smith, G. et al. Nitric oxide upregulates expression of DNA-PKcs to protect cells from DNA-damaging anti-tumour agents. Nat Cell Biol 2, 339–345 (2000). https://doi.org/10.1038/35014028

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