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The ataxia-telangiectasia related protein ATR mediates DNA-dependent phosphorylation of p53

Abstract

Levels of the tumour suppressor protein p53 are increased in response to a variety of DNA damaging agents. DNA damage-induced phosphorylation of p53 occurs at serine-15 in vivo. Phosphorylation of p53 at serine-15 leads to a stabilization of the polypeptide by inhibiting its interaction with Mdm2, a protein that targets p53 for ubiquitin-dependent degradation. However, the mechanisms by which DNA damage is signalled to p53 remain unclear. Here, we report the identification of a novel DNA-activated protein kinase that phosphorylates p53 on serine-15. Fractionation of HeLa nuclear extracts and biochemical analyses indicate that this kinase is distinct from the DNA-dependent protein kinase (DNA-PK) and corresponds to the human cell cycle checkpoint protein ATR. Immunoprecipitation studies of recombinant ATR reveal that catalytic activity of this polypeptide is required for DNA-stimulated phosphorylation of p53 on serine-15. These data suggest that ATR may function upstream of p53 in a signal transduction cascade initiated upon DNA damage and provide a biochemical assay system for ATR activity.

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Acknowledgements

We thank members of the SPJ laboratory for their advice and support. In particular, we thank Graeme Smith, John Rouse, Daniel Durocher and Susan Critchlow for helpful discussions. We also thank David Lane for advice and support. Cell lines expressing ATR derivatives were a generous gift from Stephen Friend, and cyclin A/cdk2 was a gift from Mark Jackman and Jon Pines. This work was funded by grants from the Cancer Research Campaign, the Kay Kendall Leukaemia Fund and the A-T Children's Project.

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Lakin, N., Hann, B. & Jackson, S. The ataxia-telangiectasia related protein ATR mediates DNA-dependent phosphorylation of p53. Oncogene 18, 3989–3995 (1999). https://doi.org/10.1038/sj.onc.1202973

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Keywords

  • p53
  • ATR
  • ATM
  • DNA-PK
  • ataxia telangiectasia
  • DNA damage

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