Abstract
Arylamine N-acetyltransferases (NAT) catalyze the biotransformation of many important arylamine drugs and procarcinogens. NAT can either detoxify or activate procarcinogens, complicating the manner in which these enzymes may participate in enhancing or preventing toxic responses to particular agents. Mice possess three NAT isoenzymes: Nat1, Nat2, and Nat3. Whereas Nat1 and Nat2 can efficiently acetylate many arylamines, few substrates appear to be appreciably metabolized by Nat3. We generated a Nat3 knockout mouse strain and used it along with our double Nat1/2(–/–) knockout strain to further investigate the functional role of Nat3. Nat3(–/–) mice showed normal viability and reproductive capacity. Nat3 expression was very low in wild-type animals and completely undetectable in Nat3(–/–) mice. In contrast, greatly elevated expression of Nat3 transcript was observed in Nat1/2(–/–) mice. We used a transcribed marker polymorphism approach to establish that the increased expression of Nat3 in Nat1/2(–/–) mice is a positional artifact of insertion of the phosphoglycerate kinase-neomycin resistance cassette in place of the Nat1/Nat2 gene region and upstream of the intact Nat3 gene, rather than a biological compensatory mechanism. Despite the increase in Nat3 transcript, the N-acetylation of p-aminosalicylate, sulfamethazine, 2-aminofluorene, and 4-aminobiphenyl was undetectable either in vivo or in vitro in Nat1/2(–/–) animals. In parallel, no difference was observed in the in vivo clearance or in vitro metabolism of any of these substrates between wild-type and Nat3(–/–) mice. Thus, Nat3 is unlikely to play a significant role in the N-acetylation of arylamines either in wild-type mice or in mice lacking Nat1 and Nat2 activities.
Footnotes
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This work was supported by an operating grant from the National Cancer Institute of Canada, with funding from the Canadian Cancer Society.
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Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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doi:10.1124/dmd.107.015396.
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ABBREVIATIONS: NAT, N-acetyltransferase(s); DME, drug metabolizing enzyme; 5-AS, 5-aminosalicylic acid; AF, 2-aminofluorene; ABP, 4-aminobiphenyl; ES, embryonic stem; PAS, p-aminosalicylic acid; SMZ, sulfamethazine; PGK, phosphoglycerate kinase; kb, kilobase(s); UTR, untranslated region; PCR, polymerase chain reaction; bp, base pair(s); HPLC, high-performance liquid chromatography; RT, reverse transcriptase.
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↵1 Current affiliation: Division of Clinical Pharmacology and Experimental Therapeutics, Children's Mercy Hospital and Clinics, Kansas City, Missouri
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↵2 Current affiliation: Amgen Inc., Thousand Oaks, California
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↵3 Current affiliation: Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada
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↵4 Current affiliation: London Regional Cancer Program, University of Western Ontario, London, Ontario, Canada.
- Received February 23, 2007.
- Accepted March 29, 2007.
- The American Society for Pharmacology and Experimental Therapeutics
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