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.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$189.00 per year
only $15.75 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Meredith, M., Orr, A. & Everett, R. Herpes simplex virus type 1 immediate-early protein Vmw110 binds strongly and specifically to a 135-kDa cellular protein. Virology 200, 457–469 (1994).
Everett, R. et al. A novel ubiquitin-specific protease is dyamically associted with the PML nuclear domain and binds to a herpesvirus regulatory protein. EMBO J. 16, 1519–1530 (1997).
Holowaty, M.N. et al. Protein profiling with Epstein-Barr nuclear antigen 1 reveals an interaction with the herpesvirus-associated ubiquitin-specific protease HAUSP/USP7. J. Biol. Chem. 278, 29987–29994 (2003).
Li, M. et al. Deubiquitination of p53 by HAUSP is an important pathway for p53 stabilization. Nature 416, 648–653 (2002).
Li, M., Brooks, C.L., Kon, N. & Gu, W. A dynamic role of HAUSP in the p53-Mdm2 pathway. Mol. Cell 13, 879–886 (2004).
Cummins, J.M. et al. Tumour suppression: disruption of HAUSP gene stabilizes p53. Nature 428, 486–487 (2004).
Cummins, J.M. & Vogelstein, B. HAUSP is required for p53 destabilization. Cell Cycle 3, 689–692 (2004).
Meulmeester, E. et al. Loss of HAUSP-mediated deubiquitination contributes to DNA damage-induced destabilization of Hdmx and Hdm2. Mol. Cell 18, 565–576 (2005).
Holowaty, M.N., Sheng, Y., Nguyen, T., Arrowsmith, C. & Frappier, L. Protein interaction domains of the ubiqutin specific protease, USP7/HAUSP. J. Biol. Chem. 278, 47753–47761 (2003).
Hu, M. et al. Crystal structure of a UBP-family deubiquitinating enzyme in isolation and in complex with ubiquitin aldehyde. Cell 111, 1041–1054 (2002).
Saridakis, V. et al. Structure of the p53 binding domain of HAUSP/USP7 bound to Epstein-Barr nuclear antigen 1 implications for EBV-mediated immortalization. Mol. Cell 18, 25–36 (2005).
Ye, H. et al. Distinct molecular mechanism for initiating TRAF6 signalling. Nature 418, 443–446 (2002).
Ye, H., Park, Y.C., Kreishman, M., Kieff, E. & Wu, H. The structural basis for the recognition of diverse receptor sequences by TRAF2. Mol. Cell 4, 321–330 (1999).
Rodriguez, M.S., Desterro, J.M., Lain, S., Lane, D.P. & Hay, R.T. Multiple C-terminal lysine residues target p53 for ubiquitin-proteasome-mediated degradation. Mol. Cell. Biol. 20, 8458–8467 (2000).
Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997).
Brunger, A.T. et al. Crystallography & NMR system: a new software suite for macromolecular structure determination. Acta Crystallogr. D Biol. Crystallogr. 54, 905–921 (1998).
Jones, T.A., Zou, J.Y., Cowan, S.W. & Kjeldgaard, M. Improved methods for building protein models in electron density maps and the location of errors in these models. Acta Crystallogr. A 47, 110–119 (1991).
Christopher, J.A. SPOCK (structural properties observation and calculation kit). The Center for Macromolecular Design (Texas A&M University, College Station, Texas, USA, 1998).
DeLano, W.L. The PyMOL Molecular Graphics System (DeLano Scientific, San Carlos, Californa, USA, 2002).
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.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nsmb1067
This article is cited by
-
SCML2 contributes to tumor cell resistance to DNA damage through regulating p53 and CHK1 stability
Cell Death & Differentiation (2023)
-
miR-221/222 induce instability of p53 By downregulating deubiquitinase YOD1 in acute myeloid leukemia
Cell Death Discovery (2023)
-
The roles of ubiquitination in AML
Annals of Hematology (2023)
-
USP19 Negatively Regulates p53 and Promotes Cervical Cancer Progression
Molecular Biotechnology (2023)
-
The deubiquitinase USP7 promotes HNSCC progression via deubiquitinating and stabilizing TAZ
Cell Death & Disease (2022)