title: Y-family DNA polymerase-independent gap-filling translesion synthesis across aristolochic acid-derived adenine adducts in mouse cells.
abstract: Translesion DNA synthesis (TLS) operates when replicative polymerases are blocked by DNA lesions. To investigate the mechanism of mammalian TLS, we employed a plasmid bearing a single 7-(deoxyadenosine-N6-yl)-aristolactam I (dA-AL-I) adduct, which is generated by the human carcinogen, aristolochic acid I, and genetically engineered mouse embryonic fibroblasts. This lesion induces A to T transversions at a high frequency. The simultaneous knockouts of the Polh, Poli and Polk genes did not influence the TLS efficiency or the coding property of dA-AL-I, indicating that an unknown DNA polymerase(s) can efficiently catalyze the insertion of a nucleotide opposite the adduct and subsequent extension. Similarly, knockout of the Rev1 gene did not significantly affect TLS. However, knockout of the Rev3l gene, coding for the catalytic subunit of polζ, drastically suppressed TLS and abolished dA-AL-I to T transversions. The results support the idea that Rev1 is not essential for the cellular TLS functions of polζ in mammalian cells. Furthermore, the frequency of dA-AL-I to T transversion was affected by a sequence context, suggesting that TLS, at least in part, contributes to the formation of mutational hot and cold spots observed in aristolochic acid-induced cancers.
authors: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794, United States. Electronic address: keiji.hashimoto@stonybrook.edu.
authors: Keiji
authors: K
authors: Hashimoto
authors: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794, United States.
authors: Radha
authors: R
authors: Bonala
authors: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794, United States; Department of Chemistry, Stony Brook University, Stony Brook, NY 11794, United States.
authors: Francis
authors: F
authors: Johnson
authors: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794, United States; Department of Medicine, Stony Brook University, Stony Brook, NY 11794, United States.
authors: Arthur P
authors: AP
authors: Grollman
authors: Department of Pharmacological Sciences, Stony Brook University, Stony Brook, NY 11794, United States.
authors: Masaaki
authors: M
authors: Moriya
mesh_terms: D000818:Animals; D034341:Aristolochic Acids / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001483:Base Sequence; D002273:Carcinogens / Q000494:pharmacology*; D018736:DNA Adducts / Q000096:biosynthesis* / Q000737:chemistry; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D055786:Gene Knockout Techniques; D006801:Humans; D051379:Mice; D009713:Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D000094769:DNA Polymerase iota; D000099270:Y-Family DNA Polymerases
chemical_list: D034341:Aristolochic Acids; D002273:Carcinogens; D018736:DNA Adducts; D004259:DNA-Directed DNA Polymerase; D009713:Nucleotidyltransferases; D000094769:DNA Polymerase iota; C121135:aristolactam I; C000228:aristolochic acid I; C404302:POLK protein, human; D000099270:Y-Family DNA Polymerases; C112338:Rad30 protein; C496086:Rev1 protein, mouse
keywords: Aristolochic acid; Mutational hotspot; Polζ; Rev1; Translesion DNA synthesis; Y-family polymerases