Functional polymorphisms in carboxylesterase1A2 (CES1A2) gene involves specific protein 1 (Sp1) binding sites
Section snippets
Materials and methods
GenomicDNAmaterial. We used the genomic DNA from Japanese hypertensive samples collected in the previous study [7]. These samples were given written informed consent for the use of this study. The institutional review board of the Medical Research Institute, Tokyo Medical and Dental University approved the study. Genomic DNA was extracted from whole peripheral blood using the Genomix DNA extraction kit (SRL, Tokyo, Japan).
SpecificamplificationofCES1A2genespromoter. In order to distinguish CES1A2
SNP haplotype of CES1A2 promoter
We sequenced 105 Japanese samples for the promoter regions of CES1A2, and individually investigated SNP sites. Five samples were excluded since they did not give rise to an amplified band. In the 922 bp CES1A2 promoter region, we identified a total of 10 SNPs at positions −816, −674, −427, −62, −47, −46, −41, −40, −37, and −32, and one I/D at −34 (Table 1). These sequences of CES1A2 promoters together with SNP sites were registered in the DDBJ/EMBL/GenBank databases under the Accession No. AB195643
Discussion
The current study was undertaken to clarify the molecular mechanism by which a CES1A2 polymorphism −816A/C associate with responder and non-responder of medication with a CES1-dependent ACE inhibitor [7]. We re-sequenced the CES1A2 promoter and found polymorphisms that collectively build two haplotypes: one with and another without putative Sp1 transcription factor binding sites. Transfection assay and EMSA showed that the Sp1 binding site in the promoter region is indeed functional. Since the
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