Distinct Roles of XPF-ERCC1 and Rad1-Rad10-Saw1 in Replication-Coupled and Uncoupled Inter-Strand Crosslink Repair

Nat Commun. 2018 May 23;9(1):2025. doi: 10.1038/s41467-018-04327-0.

Abstract

Yeast Rad1-Rad10 (XPF-ERCC1 in mammals) incises UV, oxidation, and cross-linking agent-induced DNA lesions, and contributes to multiple DNA repair pathways. To determine how Rad1-Rad10 catalyzes inter-strand crosslink repair (ICLR), we examined sensitivity to ICLs from yeast deleted for SAW1 and SLX4, which encode proteins that interact physically with Rad1-Rad10 and bind stalled replication forks. Saw1, Slx1, and Slx4 are critical for replication-coupled ICLR in mus81 deficient cells. Two rad1 mutations that disrupt interactions between Rpa1 and Rad1-Rad10 selectively disable non-nucleotide excision repair (NER) function, but retain UV lesion repair. Mutations in the analogous region of XPF also compromised XPF interactions with Rpa1 and Slx4, and are proficient in NER but deficient in ICLR and direct repeat recombination. We propose that Rad1-Rad10 makes distinct contributions to ICLR depending on cell cycle phase: in G1, Rad1-Rad10 removes ICL via NER, whereas in S/G2, Rad1-Rad10 facilitates NER-independent replication-coupled ICLR.

Publication types

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

MeSH terms

  • Animals
  • CHO Cells
  • Cell Cycle / genetics
  • Cricetulus
  • Cross-Linking Reagents / toxicity
  • DNA Damage / drug effects
  • DNA Damage / genetics*
  • DNA Damage / radiation effects
  • DNA Repair / physiology*
  • DNA Repair Enzymes / genetics
  • DNA Repair Enzymes / metabolism*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Endonucleases / genetics
  • Endonucleases / metabolism*
  • Intravital Microscopy
  • Mutagenesis, Site-Directed
  • Mutation
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Single-Strand Specific DNA and RNA Endonucleases / genetics
  • Single-Strand Specific DNA and RNA Endonucleases / metabolism*
  • Ultraviolet Rays / adverse effects

Substances

  • Cross-Linking Reagents
  • DNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Saw1 protein, S cerevisiae
  • xeroderma pigmentosum group F protein
  • Endonucleases
  • RAD1 protein, S cerevisiae
  • RAD10 protein, S cerevisiae
  • Single-Strand Specific DNA and RNA Endonucleases
  • DNA Repair Enzymes