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Functional effects of N‐acetyltransferase 1 (NAT1*10) polymorphisms
Author(s) -
Millner Lori Michele,
Barker David F.,
Doll Mark A.,
States J. Christopher,
Hein David W.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.lb394
Subject(s) - arylamine n acetyltransferase , chinese hamster ovary cell , polyadenylation , microbiology and biotechnology , biology , n acetyltransferase , genetics , untranslated region , acetyltransferase , messenger rna , haplotype , acetylation , gene , allele , cell culture
N ‐acetyltransferase 1 (NAT1) catalyzes N ‐acetylation of aromatic and heterocyclic amine carcinogens resulting in their activation or inactivation. NAT1*10 , a common NAT1 variant in many ethnic groups, is associated with increased risk for numerous cancers and congenital defects. NAT1*10 has putatively been described as a rapid acetylator allele (haplotype) and is characterized by two polymorphisms located in the 3' UTR, one of which disrupts a polyadenylation (polyA) signal. We determined polyA patterns of NAT1*10 compared to referent NAT1*4 using RNase protection assays (RPAs). We employed a novel approach to study functional differences caused by NAT1*10 polymorphisms by using constructs that mimic complete human mRNAs. Plasmid constructs of NAT1*10 and NAT1*4 contained full length human mRNAs including either the NATa (alternative promoter) or NATb (major promoter) 5′‐UTR, the ORF, and 885 base pairs of the 3'UTR region. Following transient transfection into Chinese hamster ovary cells, NAT1‐catalyzed N ‐acetylation of p ‐aminobenzoic acid was measured by HPLC and NAT1 protein expression was measured by Western blot. mRNA levels were determined by RT‐PCR and polyadenylation patterns by RPA. No differences were observed between NAT1*10 and NAT1*4 in levels of N ‐acetyltransferase activity, NAT1 protein and mRNA, or polyA pattern. Supported by USPHS grants CA034627, ES011564, and ES014443 and DOD BC083107.