Pirh2 E3 ubiquitin ligase targets DNA polymerase eta for 20S proteasomal degradation

Mol Cell Biol. 2010 Feb;30(4):1041-8. doi: 10.1128/MCB.01198-09. Epub 2009 Dec 14.

Abstract

DNA polymerase eta (PolH), a Y family translesion polymerase, is required for repairing UV-induced DNA damage, and loss of PolH is responsible for early onset of malignant skin cancers in patients with xeroderma pigmentosum variant (XPV), an autosomal recessive disorder. Here, we show that PolH, a target of the p53 tumor suppressor, is a short-half-life protein. We found that PolH is degraded by proteasome, which is enhanced upon UV irradiation. We also found that PolH interacts with Pirh2 E3 ligase, another target of the p53 tumor suppressor, via the polymerase-associated domain in PolH and the RING finger domain in Pirh2. In addition, we show that overexpression of Pirh2 decreases PolH protein stability, whereas knockdown of Pirh2 increases it. Interestingly, we found that PolH is recruited by Pirh2 and degraded by 20S proteasome in a ubiquitin-independent manner. Finally, we observed that Pirh2 knockdown leads to accumulation of PolH and, subsequently, enhances the survival of UV-irradiated cells. We postulate that UV irradiation promotes cancer formation in part by destabilizing PolH via Pirh2-mediated 20S proteasomal degradation.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Apoptosis / radiation effects
  • Cell Line
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Humans
  • Proteasome Endopeptidase Complex / metabolism*
  • Protein Binding
  • Ubiquitin-Protein Ligases / genetics
  • Ubiquitin-Protein Ligases / metabolism*

Substances

  • RCHY1 protein, human
  • Ubiquitin-Protein Ligases
  • DNA-Directed DNA Polymerase
  • Rad30 protein
  • Proteasome Endopeptidase Complex