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Differential promoter activities of functional haplotypes in the 5′‐flanking region of human Sulfotransferase 1A1
Author(s) -
Lin ZhongNing,
Lin YuChun,
Zhang Xuemei,
Kadlubar Susan,
Tuo Jingsheng,
Green Bridgett,
Deng Helen,
Ning Baitang
Publication year - 2012
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.21437
Subject(s) - haplotype , single nucleotide polymorphism , microbiology and biotechnology , 5' flanking region , genetics , gene , allele , biology , sulfotransferase , electrophoretic mobility shift assay , promoter , reporter gene , gene expression , genotype , sulfation
Previously, we reported five common single nucleotide polymorphisms (SNPs), −624G>C, −396G>A, −358A>C, −341C>G, and −294T>C, and six common haplotypes (CGACT, GAACT, GGAGC, GGACC, CAACT, and GAACC) in the 5′‐flanking region of the SULT1A1 gene that were associated with altered enzymatic activity. In the present study, we performed in vitro assays to determine the functional impact of these genetic variations on the promoter activity. Dual luciferase reporter assays revealed that these SNPs are located in a negative regulatory fragment of the SULT1A1 gene. Further experiments demonstrated that these SNPs and haplotypes affected promoter activities of SULT1A1 . Electrophoretic mobility shift assays showed distinctive binding patterns for the SNPs ‐396G>A and ‐294T>C, due to differential binding affinities of the G/A alleles and the T/C alleles to nuclear proteins extracted from the liver carcinoma cell lines, HepG2 and Huh7. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:422–428, 2012; View this article online at wileyonlinelibrary.com . DOI 10:1002/jbt.21437