DCAF7 is required for maintaining the cellular levels of ERCC1-XPF and nucleotide excision repair

Biochem Biophys Res Commun. 2019 Oct 29;519(1):204-210. doi: 10.1016/j.bbrc.2019.08.147. Epub 2019 Sep 4.

Abstract

The ERCC1-XPF heterodimer is a structure-specific endonuclease and plays multiple roles in various DNA repair pathways including nucleotide excision repair and also telomere maintenance. The dimer formation, which is mediated by their C-terminal helix-hairpin-helix regions, is essential for their endonuclease activity as well as the stability of each protein. However, the detailed mechanism of how a cellular level of ERCC1-XPF is regulated still remains elusive. Here, we report the identification of DDB1- and CUL4-associated factor 7 (DCAF7, also known as WDR68/HAN11) as a novel interacting protein of ERCC1-XPF by mass spectrometry after tandem purification. Immunoprecipitation experiments confirmed their interaction and suggested dominant association of DCAF7 with XPF but not ERCC1. Interestingly, siRNA-mediated knockdown of DCAF7, but not DDB1, attenuated the cellular level of ERCC1-XPF, which is partly dependent on proteasome. The depletion of TCP1α, one of components of the molecular chaperon TRiC/CCT known to interact with DCAF7 and promote its folding, also reduced ERCC1-XPF level. Finally, we show that the depletion of DCAF7 causes inefficient repair of UV-induced (6-4) photoproducts, which can be rescued by ectopic overexpression of XPF or ERCC1-XPF. Altogether, our results strongly suggest that DCAF7 is a novel regulator of ERCC1-XPF protein level and cellular nucleotide excision repair activity.

Keywords: DCAF7/WDR68/HAN11; ERCC1-XPF; Nucleotide excision repair; TRiC/CCT.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism*
  • Cell Line
  • DNA Repair*
  • DNA-Binding Proteins / metabolism*
  • Down-Regulation
  • Endonucleases / metabolism*
  • Humans
  • Proteasome Endopeptidase Complex / metabolism
  • Protein Binding
  • Protein Multimerization

Substances

  • Adaptor Proteins, Signal Transducing
  • DCAF7 protein, human
  • DNA-Binding Proteins
  • xeroderma pigmentosum group F protein
  • ERCC1 protein, human
  • Endonucleases
  • Proteasome Endopeptidase Complex