Replication-independent repair of DNA interstrand crosslinks.
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.
Institute for Cancer Genetics, Department of Genetics and Development, Columbia University, New York, NY 10032, USA.
Hannah L
HL
Williams
Max E
ME
Gottesman
Jean
J
Gautier
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002945:Cisplatin / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000494:pharmacology; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016193:G1 Phase / Q000187:drug effects / Q000235:genetics; D008239:Lysine / Q000235:genetics / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D016685:Mitomycin / Q000737:chemistry / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism; D016192:Resting Phase, Cell Cycle / Q000187:drug effects / Q000235:genetics; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism; D000099270:Y-Family DNA Polymerases
D002945:Cisplatin; D003432:Cross-Linking Reagents; D004247:DNA; D004259:DNA-Directed DNA Polymerase; D008239:Lysine; D016685:Mitomycin; D018809:Proliferating Cell Nuclear Antigen; D029867:Xenopus Proteins; C530191:Polk protein, Xenopus; D000099270:Y-Family DNA Polymerases