Adenine N3 is a main alkylation site of styrene oxide in double-stranded DNA.
Styrene 7,8-oxide (SO), a major metabolite of styrene, is classified as a probable human carcinogen. In the present work, salmon testis DNA was reacted with SO and the alkylation products were analysed after sequential depurination in neutral or acidic conditions followed by HPLC separation and UV-detection. A novel finding was that the N-3 position of adenine was the next most reactive alkylation site in double-stranded DNA, comprising 4% of the total alkylation, as compared to alkylation at the N-7 position of guanine, 93% of the total alkylation. Both alpha- and beta-products of SO were formed at these two sites. Other modified sites were N2-guanine (1.5%, alpha-isomer), 1-adenine (0.4%, both isomers) and N6-adenine (0.7%, both isomers) as well as 1-hypoxanthine (0.1%, alpha-isomer), formed by deamination of the corresponding 1-adenine adduct. The results indicated that in double-stranded DNA N-7 of guanine and N-3 of adenine account for 97% of alkylation by SO. However, these abundant adducts are not stable, the half-life of depurination in DNA for 3-substituted adenines being approximately 10 and approximately 20 h, for alpha- and beta-isomers, respectively, and 51 h for both isomers of 7-substituted guanines.
Koskinen
M
M
Department of Biosciences at Novum, Karolinska Institute, Huddinge, Sweden. mikko.koskinen@cnt.ki.se
Vodicka
P
P
Hemminki
K
K
D000225:Adenine / Q000737:chemistry* / Q000378:metabolism; D000477:Alkylating Agents / Q000737:chemistry* / Q000633:toxicity; D000478:Alkylation; D000818:Animals; D002273:Carcinogens / Q000737:chemistry / Q000633:toxicity; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry / Q000378:metabolism; D004852:Epoxy Compounds / Q000737:chemistry* / Q000633:toxicity; D006147:Guanine / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008297:Male; D012474:Salmon; D013737:Testis / Q000737:chemistry
D000477:Alkylating Agents; D002273:Carcinogens; D018736:DNA Adducts; D004852:Epoxy Compounds; D006147:Guanine; D004247:DNA; C013690:styrene oxide; D000225:Adenine