Multiple repair pathways mediate tolerance to chemotherapeutic cross-linking agents in vertebrate cells.
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.
Nojima
Kuniharu
K
Department of Radiation Genetics, Kyoto University, Japan.
Hochegger
Helfrid
H
Saberi
Alihossein
A
Fukushima
Toru
T
Kikuchi
Koji
K
Yoshimura
Michio
M
Orelli
Brian J
BJ
Bishop
Douglas K
DK
Hirano
Seiki
S
Ohzeki
Mioko
M
Ishiai
Masamichi
M
Yamamoto
Kazuhiko
K
Takata
Minoru
M
Arakawa
Hiroshi
H
Buerstedde
Jean-Marie
JM
Yamazoe
Mitsuyoshi
M
Kawamoto
Takuo
T
Araki
Kasumi
K
Takahashi
Jun A
JA
Hashimoto
Nobuo
N
Takeda
Shunichi
S
Sonoda
Eiichiro
E
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
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