title: Multiple repair pathways mediate tolerance to chemotherapeutic cross-linking agents in vertebrate cells.
abstract: Cross-linking agents that induce DNA interstrand cross-links (ICL) are widely used in anticancer chemotherapy. Yeast genetic studies show that nucleotide excision repair (NER), Rad6/Rad18-dependent postreplication repair, homologous recombination, and cell cycle checkpoint pathway are involved in ICL repair. To study the contribution of DNA damage response pathways in tolerance to cross-linking agents in vertebrates, we made a panel of gene-disrupted clones from chicken DT40 cells, each defective in a particular DNA repair or checkpoint pathway, and measured the sensitivities to cross-linking agents, including cis-diamminedichloroplatinum (II) (cisplatin), mitomycin C, and melphalan. We found that cells harboring defects in translesion DNA synthesis (TLS), Fanconi anemia complementation groups (FANC), or homologous recombination displayed marked hypersensitivity to all the cross-linking agents, whereas NER seemed to play only a minor role. This effect of replication-dependent repair pathways is distinctively different from the situation in yeast, where NER seems to play a major role in dealing with ICL. Cells deficient in Rev3, the catalytic subunit of TLS polymerase Polzeta, showed the highest sensitivity to cisplatin followed by fanc-c. Furthermore, epistasis analysis revealed that these two mutants work in the same pathway. Our genetic comprehensive study reveals a critical role for DNA repair pathways that release DNA replication block at ICLs in cellular tolerance to cross-linking agents and could be directly exploited in designing an effective chemotherapy.
authors.affiliation: Department of Radiation Genetics, Kyoto University, Japan.
authors.forename: Kuniharu
authors.initials: K
authors.lastname: Nojima
authors.forename: Helfrid
authors.initials: H
authors.lastname: Hochegger
authors.forename: Alihossein
authors.initials: A
authors.lastname: Saberi
authors.forename: Toru
authors.initials: T
authors.lastname: Fukushima
authors.forename: Koji
authors.initials: K
authors.lastname: Kikuchi
authors.forename: Michio
authors.initials: M
authors.lastname: Yoshimura
authors.forename: Brian J
authors.initials: BJ
authors.lastname: Orelli
authors.forename: Douglas K
authors.initials: DK
authors.lastname: Bishop
authors.forename: Seiki
authors.initials: S
authors.lastname: Hirano
authors.forename: Mioko
authors.initials: M
authors.lastname: Ohzeki
authors.forename: Masamichi
authors.initials: M
authors.lastname: Ishiai
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Minoru
authors.initials: M
authors.lastname: Takata
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Arakawa
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Buerstedde
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Yamazoe
authors.forename: Takuo
authors.initials: T
authors.lastname: Kawamoto
authors.forename: Kasumi
authors.initials: K
authors.lastname: Araki
authors.forename: Jun A
authors.initials: JA
authors.lastname: Takahashi
authors.forename: Nobuo
authors.initials: N
authors.lastname: Hashimoto
authors.forename: Shunichi
authors.initials: S
authors.lastname: Takeda
authors.forename: Eiichiro
authors.initials: E
authors.lastname: Sonoda
mesh_terms: D000818:Animals; D034961:Antigens, Nuclear / Q000378:metabolism; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D002645:Chickens; D002945:Cisplatin / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000502:physiology; D004843:Epistasis, Genetic; D052218:Fanconi Anemia Complementation Group C Protein / Q000235:genetics / Q000502:physiology; D015757:Genes, rev / Q000235:genetics / Q000502:physiology; D000072200:Ku Autoantigen; D008558:Melphalan / Q000494:pharmacology; D016685:Mitomycin / Q000494:pharmacology; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D011995:Recombination, Genetic / Q000187:drug effects; D015398:Signal Transduction
chemical_list: D034961:Antigens, Nuclear; D018906:Antineoplastic Agents, Alkylating; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; D052218:Fanconi Anemia Complementation Group C Protein; D019384:Nucleic Acid Synthesis Inhibitors; D016685:Mitomycin; D004247:DNA; C425929:DNA polymerase zeta; D004259:DNA-Directed DNA Polymerase; D000072200:Ku Autoantigen; D002945:Cisplatin; D008558:Melphalan