Replication-independent repair of DNA interstrand crosslinks

Mol Cell. 2012 Jul 13;47(1):140-7. doi: 10.1016/j.molcel.2012.05.001. Epub 2012 May 31.

Abstract

DNA interstrand crosslinks (ICLs) are cytotoxic lesions that covalently link opposite strands of the DNA helix and block DNA unwinding. ICLs are repaired during and outside S phase, and replication-independent ICL repair (RIR) is critical to maintain genomic integrity and to allow transcription in nondividing or slowly dividing cells. Here, we show that the Y family DNA polymerase kappa (Pol κ) is essential for RIR of a site-specific ICL lesion in Xenopus egg extracts, and that both its catalytic activity and UBZ domains are required for this function. We also demonstrate a requirement for PCNA and its modification on lysine 164. Finally, we show that Pol κ participates in ICL repair in mammalian cells, particularly in G0. Our results identify key components of the RIR pathway and begin to unravel its mechanism.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Cell Survival / drug effects
  • Cells, Cultured
  • Cisplatin / pharmacology
  • Cross-Linking Reagents / chemistry
  • Cross-Linking Reagents / pharmacology
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Damage*
  • DNA Repair*
  • DNA Replication
  • DNA-Directed DNA Polymerase / genetics
  • DNA-Directed DNA Polymerase / metabolism*
  • Embryo, Mammalian / cytology
  • Female
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism
  • G1 Phase / drug effects
  • G1 Phase / genetics
  • Lysine / genetics
  • Lysine / metabolism
  • Mice
  • Mice, Knockout
  • Mitomycin / chemistry
  • Mitomycin / pharmacology
  • Molecular Sequence Data
  • Mutation
  • Proliferating Cell Nuclear Antigen / genetics
  • Proliferating Cell Nuclear Antigen / metabolism
  • Resting Phase, Cell Cycle / drug effects
  • Resting Phase, Cell Cycle / genetics
  • Xenopus Proteins / genetics
  • Xenopus Proteins / metabolism*
  • Xenopus laevis / genetics
  • Xenopus laevis / metabolism

Substances

  • Cross-Linking Reagents
  • Proliferating Cell Nuclear Antigen
  • Xenopus Proteins
  • Mitomycin
  • DNA
  • DNA-Directed DNA Polymerase
  • Polk protein, Xenopus
  • Lysine
  • Cisplatin