Differential roles for DNA polymerases eta, zeta, and REV1 in lesion bypass of intrastrand versus interstrand DNA cross-links.
Translesion DNA synthesis (TLS) is a process whereby specialized DNA polymerases are recruited to bypass DNA lesions that would otherwise stall high-fidelity polymerases. We provide evidence that TLS across cisplatin intrastrand cross-links is performed by multiple translesion DNA polymerases. First, we determined that PCNA monoubiquitination by RAD18 is necessary for efficient bypass of cisplatin adducts by the TLS polymerases eta (Poleta), REV1, and zeta (Polzeta) based on the observations that depletion of these proteins individually leads to decreased cell survival, cell cycle arrest in S phase, and activation of the DNA damage response. Second, we showed that in addition to PCNA monoubiquitination by RAD18, the Fanconi anemia core complex is also important for recruitment of REV1 to stalled replication forks in cisplatin treated cells. Third, we present evidence that REV1 and Polzeta are uniquely associated with protection against cisplatin and mitomycin C-induced chromosomal aberrations, and both are necessary for the timely resolution of DNA double-strand breaks associated with repair of DNA interstrand cross-links. Together, our findings indicate that REV1 and Polzeta facilitate repair of interstrand cross-links independently of PCNA monoubiquitination and Poleta, whereas RAD18 plus Poleta, REV1, and Polzeta are all necessary for replicative bypass of cisplatin intrastrand DNA cross-links.
Hicks
J Kevin
JK
Department of Pharmacology, University of Michigan Medical School, Ann Arbor, Michigan 48109-2200, USA.
Chute
Colleen L
CL
Paulsen
Michelle T
MT
Ragland
Ryan L
RL
Howlett
Niall G
NG
Guéranger
Quentin
Q
Glover
Thomas W
TW
Canman
Christine E
CE
D001483:Base Sequence; D002453:Cell Cycle; D002460:Cell Line; D002869:Chromosome Aberrations; D002945:Cisplatin / Q000633:toxicity; D003432:Cross-Linking Reagents / Q000633:toxicity; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology*; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D064246:Mad2 Proteins; D016685:Mitomycin / Q000633:toxicity; D008954:Models, Biological; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D019384:Nucleic Acid Synthesis Inhibitors; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D011506:Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D044767:Ubiquitin-Protein Ligases; D000099270:Y-Family DNA Polymerases
D002945:Cisplatin; D003432:Cross-Linking Reagents; D004247:DNA; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase; D064246:Mad2 Proteins; D016685:Mitomycin; D009687:Nuclear Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D009713:Nucleotidyltransferases; D018809:Proliferating Cell Nuclear Antigen; D011506:Proteins; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases; C120552:MAD2L2 protein, human; C485954:RAD18 protein, human; C425929:DNA polymerase zeta; C429004:REV1 protein, human; D000099270:Y-Family DNA Polymerases; C118285:REV3L protein, human; C112338:Rad30 protein