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Phosphorylation of serines 15 and 37 is necessary for efficient accumulation of p53 following irradiation with UV

Oncogene volume 20, pages 1076–1084 (2001)Cite this article

Abstract

Changes in the phosphorylation state of p53 are important in increasing its half-life and potency as a transcription factor. To investigate their roles, serine residues 15 and 37 were mutated to alanines and the mutated proteins were expressed stably at low basal levels in Li-Fraumeni-derived p53-null human fibroblasts. The accumulation of p53 after DNA damage was analysed quantitatively in multiple clones. Mutation of serine 15, serine 37 or both impaired the accumulation of the protein after exposing the cells to ultraviolet radiation (50–100% increase for the mutant proteins, 500% increase for wild-type p53) but not after treatment with adriamycin. The diminished accumulation of mutant p53 protein is due to a reduction of basal HDM association. Analysis of p53-dependent transcription revealed that phosphorylation of serine 15 is required to maintain basal levels of p21 mRNA. These results provide new evidence for an important function of serine 37 phosphorylation, clearly distinguish the pathways of p53 activation in response to ultraviolet radiation or DNA damage inflicted by adriamycin, and reveal that serine 15 is crucial to support the p53-mediated basal expression of p21.

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Acknowledgements

We wish to thank Hermann Bujard (University of Heidelberg, Heidelberg, Germany) for pTO and pTA.hygro constructs, Michael Tainsky (MD Anderson Cancer Center, Houston, TX, USA) for the MDAH041 cells, Mary Ellen Perry (University of Wisconsin, Madison, WI, USA) for the HDM 2A10 antibody, and David Lane (University of Dundee, Dundee, Scotland) for wild-type p53 cDNA. Edward Mascha and Matthew Karafa provided help with the statistical analysis. LJH Bean was supported by NIH training grant 5-T32-GM08613. This project was funded by grant GM49345 from the National Institutes of Health.

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  1. Department of Molecular Biology, Lerner Research Institute, The Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, Ohio, USA

    Lora JH Bean & George R Stark

  2. Department of Genetics, Case Western Reserve University School of Medicine, 10900 Euclid Avenue, Cleveland, OH 44106, Ohio, USA

    Lora JH Bean & George R Stark

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Bean, L., Stark, G. Phosphorylation of serines 15 and 37 is necessary for efficient accumulation of p53 following irradiation with UV. Oncogene 20, 1076–1084 (2001). https://doi.org/10.1038/sj.onc.1204204

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