title: Rev1 and Polzeta influence toxicity and mutagenicity of Me-lex, a sequence selective N3-adenine methylating agent.
abstract: The relative toxicity and mutagenicity of Me-lex, which selectively generates 3-methyladenine (3-MeA), is dependent on the nature of the DNA repair background. Base excision repair (BER)-defective S. cerevisiae strains mag1 and apn1apn2 were both significantly more sensitive to Me-lex toxicity, but only the latter is significantly more prone to Me-lex-induced mutagenesis. To examine the contribution of translesion synthesis (TLS) DNA polymerases in the bypass of Me-lex-induced lesions, the REV3 and REV1 genes were independently deleted in the parental yeast strain and in different DNA repair-deficient derivatives: the nucleotide excision repair (NER)-deficient rad14, and the BER-deficient mag1 or apn1apn2 strains. The strains contained an integrated ADE2 reporter gene under control of the transcription factor p53. A centromeric yeast expression vector containing the wild-type p53 cDNA was treated in vitro with increasing concentrations of Me-lex and transformed into the different yeast strains. The toxicity of Me-lex-induced lesions was evaluated based on the plasmid transformation efficiency compared to the untreated vector, while Me-lex mutagenicity was assessed using the p53 reporter assay. In the present study, we demonstrate that disruption of Polzeta (through deletion of its catalytic subunit coded by REV3) or Rev1 (by REV1 deletion) increased Me-lex lethality and decreased Me-lex mutagenicity in both the NER-defective (rad14) and BER-defective (mag1; apn1apn2) strains. Therefore, Polzeta and Rev1 contribute to resistance of the lethal effects of Me-lex-induced lesions (3-MeA and derived AP sites) by bypassing lesions and fixing some mutations.
authors.affiliation: Department of Epidemiology and Prevention, National Cancer Research Institute (IST), L.go R. Benzi, 10, 16132 Genova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Monti
authors.forename: Yari
authors.initials: Y
authors.lastname: Ciribilli
authors.forename: Debora
authors.initials: D
authors.lastname: Russo
authors.forename: Alessandra
authors.initials: A
authors.lastname: Bisio
authors.forename: Chiara
authors.initials: C
authors.lastname: Perfumo
authors.forename: Virginia
authors.initials: V
authors.lastname: Andreotti
authors.forename: Paola
authors.initials: P
authors.lastname: Menichini
authors.forename: Alberto
authors.initials: A
authors.lastname: Inga
authors.forename: Xiaofen
authors.initials: X
authors.lastname: Huang
authors.forename: Barry
authors.initials: B
authors.lastname: Gold
authors.forename: Gilberto
authors.initials: G
authors.lastname: Fronza
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000737:chemistry; D016587:Antimutagenic Agents / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D045647:DNA Glycosylases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000187:drug effects*; D004260:DNA Repair / Q000502:physiology; D045643:DNA Repair Enzymes; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism; D009153:Mutagens / Q000633:toxicity*; D009429:Netropsin / Q000031:analogs & derivatives* / Q000633:toxicity; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D016159:Tumor Suppressor Protein p53; D000099270:Y-Family DNA Polymerases
chemical_list: D000225:Adenine; D016587:Antimutagenic Agents; D045647:DNA Glycosylases; D045643:DNA Repair Enzymes; D043603:DNA-(Apurinic or Apyrimidinic Site) Lyase; D004259:DNA-Directed DNA Polymerase; D004706:Endodeoxyribonucleases; D009153:Mutagens; D009429:Netropsin; D009713:Nucleotidyltransferases; D029701:Saccharomyces cerevisiae Proteins; D016159:Tumor Suppressor Protein p53; C072975:RAD14 protein, S cerevisiae; C110963:methyl lexitropsin; C025946:3-methyladenine; C425929:DNA polymerase zeta; C100951:REV1 protein, S cerevisiae; D000099270:Y-Family DNA Polymerases; C092602:REV3 protein, S cerevisiae; C083274:Apn1 protein, S cerevisiae; C445668:MAG1 protein, S cerevisiae; C468953:APN2 protein, S cerevisiae