Functional characterization of nucleotide polymorphisms in the coding region of N-acetyltransferase 1.
N-acetyltransferase 1 (NAT1) catalyses the activation and/or deactivation of aromatic and heterocyclic amine carcinogens. A genetic polymorphism in NAT1 is associated with an increased risk of various cancers and drug toxicities, but epidemiological investigations are severely compromised by a poor understanding of the relationship between NAT1 genotype and phenotype. Human reference NAT1*4 and 12 known human NAT1 allelic variants possessing nucleotide polymorphisms in the NAT1 coding region were cloned and expressed in yeast (Schizosaccharomyces pombe). Large reductions in N- and O-acetyltransferase catalytic activities were observed for recombinant NAT1 allozymes encoded by NAT1*14B, NAT1*15, NAT1*17, NAT1*19 and NAT1*22. Each of these alleles exhibited NAT1 protein expression levels below the limit of detection as measured by Western blot. No differences between high and low activity NAT1 alleles were observed in relative mRNA expression or relative transformation efficiency. The recombinant NAT1 17 and NAT1 22 allozymes showed reduced intrinsic stability when compared with NAT1 4. 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) N-acetylation was not catalysed by any of the NAT1 allozymes. Large differences in the metabolic activation via O-acetylation of 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-b]pyridine (N-hydroxy-PhIP) were noted for NAT1 allelic variants. The results of these studies suggest an important role for the NAT1 genetic polymorphism in metabolism of aromatic and heterocyclic amine carcinogens. Furthermore, these results suggest that low NAT1 phenotype results from NAT1 allelic variants that encode reduced expression of NAT1 and/or less-stable NAT1 protein.
Fretland
A J
AJ
Department of Pharmacology and Toxicology, University of Louisville School of Medicine, Louisville, Kentucky, USA.
Doll
M A
MA
Leff
M A
MA
Hein
D W
DW
D000123:Acetyltransferases / Q000235:genetics* / Q000378:metabolism*; D000483:Alleles; D001191:Arylamine N-Acetyltransferase; D002273:Carcinogens / Q000378:metabolism; D003001:Cloning, Molecular; D004795:Enzyme Stability; D014644:Genetic Variation; D006358:Hot Temperature; D006801:Humans; D007093:Imidazoles / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D011110:Polymorphism, Genetic; D011489:Protein Denaturation; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA
D002273:Carcinogens; D007093:Imidazoles; D007527:Isoenzymes; D011994:Recombinant Proteins; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D000123:Acetyltransferases; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1