Differential promoter activities of functional haplotypes in the 5'-flanking region of human sulfotransferase 1A1.
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.
Division of Personalized Nutrition and Medicine, National Center for Toxicological Research, Food and Drug Administration, Jefferson, AR 72079, USA.
Zhong-Ning
ZN
Lin
Yu-Chun
YC
Lin
Xuemei
X
Zhang
Susan
S
Kadlubar
Jingsheng
J
Tuo
Bridgett
B
Green
Helen
H
Deng
Baitang
B
Ning
D024506:5' Flanking Region; D015239:Arylsulfotransferase / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D017930:Genes, Reporter; D056726:Genetic Association Studies; D006239:Haplotypes; D056945:Hep G2 Cells; D006801:Humans; D049410:Luciferases, Renilla / Q000096:biosynthesis / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D015533:Transcriptional Activation
D049410:Luciferases, Renilla; D015239:Arylsulfotransferase; C117544:SULT1A1 protein, human