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Molecular recognition of p53 and MDM2 by USP7/HAUSP

Abstract

The ubiquitin-specific protease, USP7, has key roles in the p53 pathway whereby it stabilizes both p53 and MDM2. We show that the N-terminal domain of USP7 binds two closely spaced 4-residue sites in both p53 and MDM2, falling between p53 residues 359–367 and MDM2 residues 147–159. Cocrystal structures with USP7 were determined for both p53 peptides and for one MDM2 peptide. These peptides bind the same surface of USP7 as Epstein-Barr nuclear antigen-1, explaining the competitive nature of the interactions. The structures and mutagenesis data indicate a preference for a P/AXXS motif in peptides that bind USP7. Contacts made by serine are identical and crucial for all peptides, and Trp165 in the peptide-binding pocket of USP7 is also crucial. These results help to elucidate the mechanism of substrate recognition by USP7 and the regulation of the p53 pathway.

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Figure 1: Crystal structure of USP7 NTD bound to p53 site 359-PGGS-362.
Figure 2: Crystal structure of USP7 NTD bound to p53 site 364-AHSS-367.
Figure 3: Mapping of the MDM2-USP7 interaction.
Figure 4: Crystal structure of USP7 NTD bound to MDM2 peptide 4.
Figure 5: Peptide interactions with USP7 NTD mutants.

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Acknowledgements

We thank A. Davidson for use of his spectrofluorometer and E.F. Pai and the Structural Genomics Consortium for use of their diffractometers. This work was funded by the Canadian Cancer Society through grants to L.F. and C.H.A. from the National Cancer Institute of Canada. V.S. was supported by a Natural Sciences and Engineering Research Council of Canada post-doctoral fellowship and Y.S. was supported by a Terry Fox Foundation research fellowship from the National Cancer Institute of Canada.

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Correspondence to Lori Frappier.

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Sheng, Y., Saridakis, V., Sarkari, F. et al. Molecular recognition of p53 and MDM2 by USP7/HAUSP. Nat Struct Mol Biol 13, 285–291 (2006). https://doi.org/10.1038/nsmb1067

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