title: Mutagenic specificity of 2-acetylaminonaphthalene-derived DNA adduct in mammalian cells.
abstract: 2-Acetylaminonaphthalene (2-AAN) has been recognized as a urinary bladder carcinogen in humans. The deacetylated form, 2-aminonaphthalene (2-AN), is metabolized in vivo and reacts primarily with guanine residues in DNA, resulting in the formation of dG-N(2)-aminonaphthalene (dG-N(2)-AN) adduct. Phosphoramidite chemical procedure has recently been established in our laboratory to prepare oligodeoxynucleotides containing a single dG-N(2)-acetylaminonaphthalene (dG-N(2)-AAN) adduct. Oligodeoxynucleotides ((5')TCCTCCTNXCCTCTC, where X is dG or dG-N(2)-AAN and N is C, A, T or G) with different bases 5' flanking to the lesion were prepared and were inserted into a single-strand shuttle vectors and used to establish the mutational frequency and specificity of dG-N(2)-AAN adduct in simian kidney cells. dG-N(2)-AAN adduct promoted preferential incorporation of dCMP, the correct base, opposite the lesion. When the 5' flanking base to the lesion was C, A or T, the mutational frequency was under 2.1%. When G flanked to the lesion, the mutational frequency was slightly increased to 4.2%. Misincorporation of dAMP, dTMP, and/or dGMP varied depending on the 5' flanking base. When dG-N(2)-AAN was positioned at codon 61 of noncoding strand of human c-Ha-ras1 gene ((5')TCCTCCTXGCCTCTC, where X is dG-N(2)-AAN), the mutational frequency was 6.7%; G-->T transversions (4.7%), followed by G-->A transition (2.0%), were observed. These results demonstrated that dG-N(2)-AAN is a weak mutagenic lesion in mammalian cells. The influence of 5' flanking sequence context was observed on the mutational frequency and specificity of this adduct.
authors.affiliation: Laboratory of Chemical Biology, Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY 11794-8651, USA.
authors.forename: Xingzhi
authors.initials: X
authors.lastname: Tan
authors.forename: Radha-Rani
authors.initials: RR
authors.lastname: Bonala
authors.forename: Naomi
authors.initials: N
authors.lastname: Suzuki
authors.forename: Francis
authors.initials: F
authors.lastname: Johnson
authors.forename: Arthur P
authors.initials: AP
authors.lastname: Grollman
authors.forename: Shinya
authors.initials: S
authors.lastname: Shibutani
mesh_terms: D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000633:toxicity*; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002273:Carcinogens / Q000378:metabolism / Q000633:toxicity*; D002522:Chlorocebus aethiops; D018736:DNA Adducts / Q000235:genetics / Q000378:metabolism*; D003849:Deoxyguanosine / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism
chemical_list: D002273:Carcinogens; D018736:DNA Adducts; D009838:Oligodeoxyribonucleotides; C003914:2-acetylaminonaphthalene; D015081:2-Naphthylamine; D003849:Deoxyguanosine