PHD3 Regulates p53 Protein Stability by Hydroxylating Proline 359

Cell Rep. 2018 Jul 31;24(5):1316-1329. doi: 10.1016/j.celrep.2018.06.108.

Abstract

Cellular p53 protein levels are regulated by a ubiquitination/de-ubiquitination cycle that can target the protein for proteasomal destruction. The ubiquitination reaction is catalyzed by a multitude of ligases, whereas the removal of ubiquitin chains is mediated by two deubiquitinating enzymes (DUBs), USP7 (HAUSP) and USP10. Here, we show that PHD3 hydroxylates p53 at proline 359, a residue that is in the p53-DUB binding domain. Hydroxylation of p53 upon proline 359 regulates its interaction with USP7 and USP10, and its inhibition decreases the association of p53 with USP7/USP10, increases p53 ubiquitination, and rapidly reduces p53 protein levels independently of mRNA expression. Our results show that p53 is a PHD3 substrate and that hydroxylation by PHD3 regulates p53 protein stability through modulation of ubiquitination.

Keywords: EglN3; PHD3; USP7; hydroxylases; hypoxia; p53; proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • HEK293 Cells
  • Humans
  • Hypoxia-Inducible Factor-Proline Dioxygenases / metabolism*
  • Protein Binding
  • Protein Stability
  • Tumor Suppressor Protein p53 / chemistry
  • Tumor Suppressor Protein p53 / metabolism*
  • Ubiquitin Thiolesterase / metabolism
  • Ubiquitin-Specific Peptidase 7 / metabolism
  • Ubiquitination*

Substances

  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • USP10 protein, human
  • EGLN3 protein, human
  • Hypoxia-Inducible Factor-Proline Dioxygenases
  • USP7 protein, human
  • Ubiquitin Thiolesterase
  • Ubiquitin-Specific Peptidase 7