title: The benzene metabolite p-benzoquinone forms adducts with DNA bases that are excised by a repair activity from human cells that differs from an ethenoadenine glycosylase.
abstract: Benzene is a ubitiquous human environment mental carcinogen. One of the major metabolites is hydroquinone, which is oxidized in vivo to give p-benzoquinone (p-BQ). Both metabolites are toxic to human cells. p-BQ reacts with DNA to form benzetheno adducts with deoxycytidine, deoxyadenosine, and deoxyguanosine. In this study we have synthesized the exocyclic compounds 3-hydroxy-3-N4-benzetheno-2'-deoxycytidine (p-BQ-dCyd) and 9-hydroxy-1,N6-benzetheno-2'-deoxyadenosine (p-BQ-dAdo), respectively, by reacting deoxycytidine and deoxyadenosine with p-BQ. These were converted to the phosphoamidites, which were then used to prepare site-specific oligonucleotides with either the p-BQ-dCyd or p-BQ-dAdo adduct (pbqC or pbqA in sequences) at two different defined positions. These oligonucleotides were efficiently nicked 5' to the adduct by partially purified HeLa cell extracts--the pbqC-containing oligomer more rapidly than the pbqA-containing oligomer. In contrast to the enzyme binding to derivatives produced by the vinyl chloride metabolite chloroacetaldehyde, the oligonucleotides up to 60-mer containing p-BQ adducts did not bind measurably to the same enzyme preparation in a gel retardation assay. Furthermore, there was no competition for the binding observed between oligonucleotides containing 1,N6-etheno A deoxyadenosine (1,N6-etheno-dAdo; epsilon A in sequences) and these oligomers containing either of the p-BQ adducts, even at 120-fold excess. When highly purified fast protein liquid chromatography (FPLC) enzyme fractions were obtained, there appeared to be two closely eluting nicking activities. One of these enzymes bound and cleaved the epsilon A-containing deoxyoligonucleotide. The other enzyme cleaved the pbqA- and pbqC-containing deoxyoligonucleotides. One additional unexpected fact was that bulk p-BQ-treated salmon sperm DNA did compete effectively with the epsilon A-containing oligonucleotide for protein binding. This raises the possibility that such DNA contains other, as-yet-uncharacterized adducts that are recognized by the same enzyme that recognizes the etheno adducts. In summary, we describe a previously undescribed human DNA repair activity, possibly a glycosylase, that excises from DNA pbqC and pbqA, exocyclic adducts resulting from reaction of deoxycytidine and deoxyadenosine with the benzene metabolite, p-BQ. This glycosylase activity is not identical to the one previously reported from this laboratory as excising the four etheno bases from DNA.
authors.affiliation: Life Sciences Division, Lawrence Berkeley Laboratory, University of California, Berkeley 94720, USA.
authors.forename: A
authors.initials: A
authors.lastname: Chenna
authors.forename: B
authors.initials: B
authors.lastname: Hang
authors.forename: B
authors.initials: B
authors.lastname: Rydberg
authors.forename: E
authors.initials: E
authors.lastname: Kim
authors.forename: K
authors.initials: K
authors.lastname: Pongracz
authors.forename: W J
authors.initials: WJ
authors.lastname: Bodell
authors.forename: B
authors.initials: B
authors.lastname: Singer
mesh_terms: D001345:Autoradiography; D001483:Base Sequence; D016227:Benzoquinones / Q000302:isolation & purification / Q000378:metabolism* / Q000633:toxicity; D002474:Cell-Free System; D002851:Chromatography, High Pressure Liquid; D018736:DNA Adducts; D004260:DNA Repair; D003839:Deoxyadenosines; D003841:Deoxycytidine; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008969:Molecular Sequence Data; D009699:N-Glycosyl Hydrolases / Q000378:metabolism*; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000302:isolation & purification; D010761:Phosphorus Radioisotopes; D011485:Protein Binding
chemical_list: D016227:Benzoquinones; D018736:DNA Adducts; D003839:Deoxyadenosines; D009838:Oligodeoxyribonucleotides; D010761:Phosphorus Radioisotopes; D003841:Deoxycytidine; C004532:quinone; D009699:N-Glycosyl Hydrolases