title: The contribution of UDP-glucuronosyltransferase 1A9 on CYP1A2-mediated genotoxicity by aromatic and heterocyclic amines.
abstract: The importance of environmental and dietary arylamines, and heterocyclic amines in the etiology of human cancer is of growing interest. These pre-carcinogens are known to undergo bioactivation by cytochrome P450 (CYP)-directed oxidation, which then become substrates for the UDP-glucuronosyltransferases (UGTs). Thus, glucuronidation may contribute to the elimination of CYP-mediated reactive intermediate metabolites, preventing a toxic event. In this study, human UGTs were analyzed for their ability to modulate the mutagenic actions of N-hydroxy-arylamines formed by CYP1A2. Studies with recombinant human UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, UGT1A10, UGT2B4, UGT2B7 and UGT2B15 expressed in heterologous cell culture confirmed that UGT1A9 glucuronidated the mutagenic arylamines N-hydroxy-2-acetylaminofluorene (N-hydroxy-2AAF) and 2-hydroxyamino-1-methyl-6-phenylimidazo(4,5-b)pyridine (N-hydroxy-PhIP). To examine the mutagenic potential of these agents, a genotoxicity assay was employed using Salmonella typhimurium NM2009, a bacterial strain expressing the umuC SOS response gene fused to a beta-galactosidase reporter lacZ gene. DNA modification results in the induction of the umuC gene and subsequent enhancement of beta-galactosidase activity. Both N-hydroxy-2AAF and N-hydroxy-PhIP stimulated a dose-dependent increase in bacterial beta-galactosidase activity. In addition, the procarcinogens 2AAF and PhIP were efficiently bioactivated to bacterial mutagens when incubated with Escherichia coli membranes expressing CYP1A2 and NADPH reductase. CYP1A2 generated 2AAF- and PhIP-mediated DNA damage, but only the action of N-hydroxy-2AAF was blocked by expressed UGT1A9. These results indicate that UGT1A9 can control the outcome of a genotoxic response. The results also indicate that while a potential toxicant such as N-hydroxy-PhIP can serve as substrate for glucuronidation, its biological actions can exceed the capacity of the detoxification pathway to prevent the mutagenic episode.
authors.affiliation: Departments of Chemistry, Biochemistry and Pharmacology, University of California, San Diego, La Jolla, CA 92093-0636, USA.
authors.forename: M F
authors.initials: MF
authors.lastname: Yueh
authors.forename: N
authors.initials: N
authors.lastname: Nguyen
authors.forename: M
authors.initials: M
authors.lastname: Famourzadeh
authors.forename: C P
authors.initials: CP
authors.lastname: Strassburg
authors.forename: Y
authors.initials: Y
authors.lastname: Oda
authors.forename: F P
authors.initials: FP
authors.lastname: Guengerich
authors.forename: R H
authors.initials: RH
authors.lastname: Tukey
mesh_terms: D015073:2-Acetylaminofluorene / Q000493:pharmacokinetics / Q000633:toxicity; D000818:Animals; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D001711:Biotransformation; D002384:Catalysis; D002460:Cell Line; D019388:Cytochrome P-450 CYP1A2 / Q000235:genetics / Q000378:metabolism*; D004259:DNA-Directed DNA Polymerase; D029968:Escherichia coli Proteins; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D020719:Glucuronides / Q000096:biosynthesis; D014453:Glucuronosyltransferase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006881:Hydroxyacetylaminofluorene / Q000493:pharmacokinetics* / Q000633:toxicity*; D006900:Hydroxylation; D007093:Imidazoles / Q000493:pharmacokinetics* / Q000633:toxicity*; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D051379:Mice; D009152:Mutagenicity Tests; D009153:Mutagens / Q000493:pharmacokinetics* / Q000633:toxicity*; D011725:Pyridines / Q000493:pharmacokinetics* / Q000633:toxicity*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D020719:Glucuronides; D007093:Imidazoles; D007527:Isoenzymes; D009153:Mutagens; D011725:Pyridines; D011994:Recombinant Proteins; C060533:2-hydroxyamino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D006881:Hydroxyacetylaminofluorene; C078202:UmuC protein, E coli; D015073:2-Acetylaminofluorene; D019388:Cytochrome P-450 CYP1A2; D014453:Glucuronosyltransferase; D004259:DNA-Directed DNA Polymerase