DNA damage induced by the environmental carcinogen butadiene: identification of a diepoxybutane-adenine adduct and its detection by 32P-postlabelling.
To date only a few studies have been undertaken on DNA adducts formed by epoxybutene (EB) and diepoxybutane (DEB), the two active metabolites of 1,3-butadiene. Our interests have focused on further investigating DNA alkylation by the two epoxides, especially in relation to the development of a method for human biomonitoring. Here, following the reaction of deoxyadenosine monophosphate and poly(dA-dT)(dA-dT) with DEB and subsequent HPLC, we have identified an adenine adduct. MS analyses indicate the structure of an adenine adducted by DEB at the N6 position. A HPLC/32P-postlabelling method was developed for its measurement in DNA samples and the adduct was detected in calf thymus DNA and DNA from Chinese hamster ovary cells exposed to DEB. The 100% labelling efficiency during postlabelling, the amount of the adduct and its elution before the normal nucleotides during HPLC suggest it could be a suitable indicator of BUT exposure.
Leuratti
C
C
Molecular Biology Research Group, School of Biological Sciences, University College of Swansea, UK.
Jones
N J
NJ
Marafante
E
E
Kostiainen
R
R
Peltonen
K
K
Waters
R
R
D000218:Adamantane / Q000031:analogs & derivatives / Q000378:metabolism; D000225:Adenine / Q000032:analysis* / Q000378:metabolism*; D000818:Animals; D002070:Butadienes / Q000378:metabolism* / Q000633:toxicity*; D016466:CHO Cells; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004247:DNA / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism; D018736:DNA Adducts / Q000032:analysis* / Q000302:isolation & purification / Q000378:metabolism*; D004249:DNA Damage; D004785:Environmental Pollutants / Q000378:metabolism* / Q000633:toxicity*; D004852:Epoxy Compounds / Q000032:analysis* / Q000378:metabolism* / Q000633:toxicity; D007553:Isotope Labeling / Q000379:methods; D010761:Phosphorus Radioisotopes; D011067:Poly dA-dT / Q000378:metabolism; D016339:Spectrometry, Mass, Fast Atom Bombardment
D002070:Butadienes; D018736:DNA Adducts; D004785:Environmental Pollutants; D004852:Epoxy Compounds; D010761:Phosphorus Radioisotopes; D011067:Poly dA-dT; C016358:2,4-diamino-5-adamantyl-6-methylpyrimidine; C007366:diepoxybutane; D004247:DNA; D000225:Adenine; D000218:Adamantane