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Regulation by phosphorylation of the relative affinities of the N-terminal transactivation domains of p53 for p300 domains and Mdm2

Oncogene volume 28, pages 2112–2118 (2009)Cite this article

Abstract

The transcriptional activity of the tumour suppressor, p53, requires direct binding between its transactivation domain (TAD, 1–57) and the transcriptional coactivator, p300. We systematically assessed the role of TAD phosphorylation on binding of the p300 domains CH3, Taz1, Kix and IBiD. Thr18 phosphorylation increased the affinity up to sevenfold for CH3 and Taz1, with smaller increases from phosphorylation of Ser20, Ser15, Ser37, Ser33, Ser46 and Thr55. Binding of Kix and IBiD was less sensitive to phosphorylation. Strikingly, hepta-phosphorylation of all Ser and Thr residues increased binding 40- and 80-fold with CH3 and Taz1, respectively, but not with Kix or IBiD. Substitution of all phospho-sites with aspartates partially mimicked the effects of hepta-phosphorylation. Mdm2, the main negative regulator of p53, competes with p300 for binding to TAD. Binding of Mdm2 to TAD was reduced significantly only on phosphorylation of Thr18 (sevenfold) or by hepta-phosphorylation (24-fold). The relative affinities of Mdm2 and p300 for p53 TAD can thus be changed by up to three orders of magnitude by phosphorylation. Accordingly, phosphorylation of Thr18 and hepta-phosphorylation dramatically shifts the balance towards favouring the binding of p300 with p53, and is thus likely to be an important factor in its regulation.

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Abbreviations

CH1/CH3:

cysteine-histidine-rich regions 1 and 3

TAD:

p53 transactivation domain

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Acknowledgements

We thank Drs Jenifer Lum and Hadassah Sade for helpful suggestions and critical reading of the paper.

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  1. Department of Chemistry, MRC Centre for Protein Engineering and Cambridge University, MRC Centre, Cambridge, UK

    D P Teufel, M Bycroft & A R Fersht

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  1. D P Teufel
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Correspondence to A R Fersht.

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Teufel, D., Bycroft, M. & Fersht, A. Regulation by phosphorylation of the relative affinities of the N-terminal transactivation domains of p53 for p300 domains and Mdm2. Oncogene 28, 2112–2118 (2009). https://doi.org/10.1038/onc.2009.71

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