QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: dmd.107.016485v1 35/9/1649    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Husain, A. Articles by Hein, D. W. Search for Related Content PubMed PubMed Citation Articles by Husain, A. Articles by Hein, D. W.

Functional Analysis of the Human N-Acetyltransferase 1 Major Promoter: Quantitation of Tissue Expression and Identification of Critical Sequence Elements

Department of Pharmacology & Toxicology, James Graham Brown Cancer Center and Center for Environmental Genomics & Integrative Biology, University of Louisville School of Medicine, Louisville, Kentucky

Arylamine N-acetyltransferase 1 (NAT1) plays an important role in the biotransformation of xenobiotics, and genetic variants have been implicated in susceptibility to cancer and birth defects. A specific and quantitative reverse transcription-polymerase chain reaction assay for transcription from the major NAT1 promoter detected high expression with limited variability in human tissues. A 213-base pair (bp) minimal promoter was identified by transfection of luciferase reporter constructs into MCF-7 and HepG2 cell lines. Alignment of the 213-bp region with paralogous and orthologous promoters revealed two conserved region segments, one of which overlaps a 16-bp perfect palindrome. Transfection of luciferase constructs with artificial mutations in the minimal promoter defined two sites important for promoter function. One of these sites included a close match to the Sp1 transcription factor binding consensus sequence. Electrophoretic mobility shift assays (EMSAs), followed by competitive and supershift analyses, confirmed the Sp1 binding. Mutation of the highly conserved portion of the 16-bp palindrome reduced promoter activity more than 3-fold, and an EMSA shift was detected with an oligonucleotide, 200L29, which spans this segment. The 200L29 EMSA shift could not be competed by consensus Sp1 or AP-2 oligonucleotides, and may represent binding of a transcription factor that is common to N-acetyltransferase genes in humans and other species.

This article has been cited by other articles:

				F. A. Jefferson, G. H. Xiao, and D. W. Hein 4-Aminobiphenyl Downregulation of NAT2 Acetylator Genotype-Dependent N- and O-acetylation of Aromatic and Heterocyclic Amine Carcinogens in Primary Mammary Epithelial Cell Cultures from Rapid and Slow Acetylator Rats Toxicol. Sci., January 1, 2009; 107(1): 293 - 297. [Abstract] [Full Text] [PDF]

				D. F. Barker, J. M. Walraven, E. H. Ristagno, M. A. Doll, J. C. States, and D. W. Hein Quantitative Tissue and Gene-Specific Differences and Developmental Changes in Nat1, Nat2, and Nat3 mRNA Expression in the Rat Drug Metab. Dispos., December 1, 2008; 36(12): 2445 - 2451. [Abstract] [Full Text] [PDF]

				D. W. Hein, J. Bendaly, J. R. Neale, and M. A. Doll Systemic Functional Expression of N-Acetyltransferase Polymorphism in the F344 Nat2 Congenic Rat Drug Metab. Dispos., December 1, 2008; 36(12): 2452 - 2459. [Abstract] [Full Text] [PDF]