Regulation of the expression of human organic anion transporter 3 by hepatocyte nuclear factor 1α/β and DNA methylation

Ryota Kikuchi, Hiroyuki Kusuhara, Naka Hattori, Kunio Shiota, Insook Kim, Frank J. Gonzalez, Yuichi Sugiyama

Research output: Contribution to journalArticle

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Abstract

Human organic anion transporter 3 (hOAT3/SLC22A8) is predominantly expressed in the proximal tubules of the kidney and plays a major role in the urinary excretion of a variety of organic anions. The promoter region of hOAT3 was characterized to elucidate the mechanism underlying the tissue-specific expression of hOAT3. The minimal promoter of hOAT3 was identified to be located approximately 300 base pairs upstream of the transcriptional start site, where there are canonical TATA and hepatocyte nuclear factor (HNF1) binding motifs, which are conserved in the rodent Oat3 genes. Transactivation assays revealed that HNF1α and HNF1β markedly increased hOAT3 promoter activity, where the transactivation potency of HNF1β was lower than that of HNF1α. Mutations in the HNF1 binding motif prevented the transactivation. Electrophoretic mobility shift assays demonstrated binding of the HNF1α/HNF1α homodimer or HNF1α/HNF1β heterodimer to the hOAT3 promoter. It was also demonstrated that the promoter activity of hOAT3 is repressed by DNA methylation. Moreover, the expression of hOAT3 was activated de novo by forced expression of HNF1α alone or both HNF1α and HNF1β together with the concomitant DNA demethylation in human embryonic kidney 293 cells that lack expression of endogenous HNF1β and HNF1β, whereas forced expression of HNF1β alone could not activate the expression of hOAT3. This suggests a synergistic action of the HNF1α/HNF1α homodimer or HNF1α/HNF1β heterodimer and DNA demethylation for the constitutive expression of hOAT3. These results indicate that the tissue-specific expression of hOAT3 might be regulated by the concerted effect of genetic (HNF1α and HNF1β) and epigenetic (DNA methylation) factors.

Original languageEnglish
Pages (from-to)887-896
Number of pages10
JournalMolecular Pharmacology
Volume70
Issue number3
DOIs
Publication statusPublished - 2006
Externally publishedYes

Fingerprint

Hepatocyte Nuclear Factor 1
Organic Anion Transporters
DNA Methylation
Transcriptional Activation
Hepatocyte Nuclear Factors
Proximal Kidney Tubule
DNA
Electrophoretic Mobility Shift Assay
Genetic Promoter Regions
Epigenomics
Base Pairing
Anions
Rodentia
Kidney
Mutation
Genes

ASJC Scopus subject areas

  • Pharmacology

Cite this

Kikuchi, R., Kusuhara, H., Hattori, N., Shiota, K., Kim, I., Gonzalez, F. J., & Sugiyama, Y. (2006). Regulation of the expression of human organic anion transporter 3 by hepatocyte nuclear factor 1α/β and DNA methylation. Molecular Pharmacology, 70(3), 887-896. https://doi.org/10.1124/mol.106.025494

Regulation of the expression of human organic anion transporter 3 by hepatocyte nuclear factor 1α/β and DNA methylation. / Kikuchi, Ryota; Kusuhara, Hiroyuki; Hattori, Naka; Shiota, Kunio; Kim, Insook; Gonzalez, Frank J.; Sugiyama, Yuichi.

In: Molecular Pharmacology, Vol. 70, No. 3, 2006, p. 887-896.

Research output: Contribution to journalArticle

Kikuchi, R, Kusuhara, H, Hattori, N, Shiota, K, Kim, I, Gonzalez, FJ & Sugiyama, Y 2006, 'Regulation of the expression of human organic anion transporter 3 by hepatocyte nuclear factor 1α/β and DNA methylation', Molecular Pharmacology, vol. 70, no. 3, pp. 887-896. https://doi.org/10.1124/mol.106.025494
Kikuchi, Ryota ; Kusuhara, Hiroyuki ; Hattori, Naka ; Shiota, Kunio ; Kim, Insook ; Gonzalez, Frank J. ; Sugiyama, Yuichi. / Regulation of the expression of human organic anion transporter 3 by hepatocyte nuclear factor 1α/β and DNA methylation. In: Molecular Pharmacology. 2006 ; Vol. 70, No. 3. pp. 887-896.
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