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N-acetyltransferase 1 (NAT1) catalyzesN-acetylation of arylamines as well as theO-acetylation ofN-hydroxylated arylamines.O-acetylation leads to the formation of electrophilic intermediates that result in DNA adducts and mutations.NAT1*10is the most common variant haplotype and is associated with increased risk for numerous cancers. NAT1 is transcribed from a major promoter, NATb, and an alternative promoter, NATa, resulting in messenger RNAs (mRNAs) with distinct 5′-untranslated regions (UTRs). To best mimicin vivometabolism and the effect ofNAT1*10polymorphisms on polyadenylation usage, pcDNA5/Flp recombination target plasmid constructs were prepared for transfection of full-length human mRNAs including the 5′-UTR derived from NATb, the open reading frame and 888 nucleotides of the 3′-UTR. Following stable transfection ofNAT1*4,NAT1*10and an additionalNAT1*10variant (termedNAT1*10B) into nucleotide excision repair-deficient Chinese hamster ovary cells,N- andO-acetyltransferase activity (in vitroandin situ), mRNA and protein expression were higher in cells transfected withNAT1*10andNAT1*10Bthan in cells transfected withNAT1*4(P&lt; 0.05). Consistent with NAT1 expression and activity, cytotoxicity and hypoxanthine phosphoribosyl transferase mutants following 4-aminobiphenyl exposures were higher inNAT1*10than inNAT1*4transfected cells. Ribonuclease protection assays showed no difference betweenNAT1*4andNAT1*10. However, protection of one probe byNAT1*10Bwas not observed withNAT1*4orNAT1*10, suggesting additional mechanisms that regulateNAT1*10B. The higher mutants in cells transfected withNAT1*10andNAT1*10Bare consistent with an increased cancer risk for individuals possessingNAT1*10haplotypes.

carcinogenesis

Functional analysis of arylamine               N              -acetyltransferase 1 (              NAT1              )               NAT1*10               haplotypes in a complete NATb mRNA construct