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p53-Dependent growth arrest and altered p53-immunoreactivity following metabolic labelling with 32P ortho-phosphate in human fibroblasts

Oncogene volume 18, pages 3788–3792 (1999)Cite this article

Abstract

The tumour suppressor gene p53 plays a major role in the cellular response to DNA damage, mediating growth arrest and/or apoptosis. Phosphorylation of the protein occurs at numerous sites in vivo and is likely to be a major mechanism for modulation of its activity as a transcriptional transactivator. Not surprisingly, therefore, p53 has been intensively studied by 32P metabolic labelling. Here we show however, using normal human fibroblasts, that typical labelling conditions induce (i) a p53-dependent inhibition of DNA synthesis and (ii) an increase in the cellular content of p53 protein detectable by the phosphorylation-sensitive antibody DO-1 but not by antibody DO-12. These data demonstrate for the first time that 32P labelling is sufficient to induce a biologically-significant, p53-mediated cellular response and strongly suggest that it perturbs the phosphorylation state of p53 which it is being used to measure. This highlights the need to re-evaluate earlier data by non-radioactive approaches using phospho-specific antibodies.

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Acknowledgements

We are grateful to the Cancer Research Campaign for grant support, and to Theresa King for manuscript preparation.

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Authors and Affiliations

  1. Department of Pathology, Cancer Research Campaign Laboratories, University of Wales College of Medicine, CF4 4XN, Cardiff, UK

    Jane A Bond, Katherine Webley, Fiona S Wyllie, Christopher J Jones & David Wynford-Thomas

  2. Department of Molecular and Cellular Pathology, Ninewells Hospital Medical School, University of Dundee, Dundee, DD1 9SY, UK

    Ashley Craig & Ted Hupp

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  1. Jane A Bond
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Bond, J., Webley, K., Wyllie, F. et al. p53-Dependent growth arrest and altered p53-immunoreactivity following metabolic labelling with 32P ortho-phosphate in human fibroblasts. Oncogene 18, 3788–3792 (1999). https://doi.org/10.1038/sj.onc.1202733

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