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Identification of regulatory sites in the human PXR (NR1I2) promoter region

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Abstract

The human pregnane X receptor (hPXR, NR1I2) is a member of the nuclear receptor superfamily and a key regulator of genes encoding several major cytochrome P450 enzymes and transporters. However, the transcriptional regulation of hPXR itself remains unclear. We recently reported significant diversity in the 5′ region of human hepatic PXR transcripts and identified the major transcription initiation site. Here, we investigate the transcriptional regulatory sites in the hPXR 5′-flanking region. Luciferase reporter constructs containing various lengths of 5′-flanking region, up to 10.5 kb upstream of the major transcription initiation site, were assessed for promoter activity in HepG2 cells. We mapped the minimal essential region for promoter activity to a 160 bp region upstream of the transcription initiation site, an area that also showed nuclear protein binding. Constructs with mutations introduced into these protein-binding sites demonstrated reduced promoter activity concomitant with reduced DNA-protein binding activity. hPXR promoter activity was observed in HepG2 cells but not in HeLa cells. Likewise, nuclear protein binding to promoter elements was also observed in HepG2 but not HeLa cells. The present study provides basic information on the transcriptional regulation of hPXR and may help elucidate the regulatory mechanisms of hPXR target genes.

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Authors and Affiliations

  1. Project Team for Pharmacogenetics, National Institute of Health Sciences, Tokyo, Japan

    Kouichi Kurose, Shinobu Ikeda, Satoru Koyano & Jun-ichi Sawada

  2. Division of Medicinal Safety Science, National Institute of Health Sciences, Tokyo, Japan

    Kouichi Kurose, Masahiro Tohkin & Ryuichi Hasegawa

  3. Division of Biochemistry and Immunochemistry, National Institute of Health Sciences, Kamiyoga, Setagaya-ku, Tokyo, Japan

    Jun-ichi Sawada

  4. Division of Medicinal Safety Science, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan

    Kouichi Kurose

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  1. Kouichi Kurose
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  2. Shinobu Ikeda
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  3. Satoru Koyano
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  4. Masahiro Tohkin
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  5. Ryuichi Hasegawa
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  6. Jun-ichi Sawada
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Correspondence to Kouichi Kurose.

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Kurose, K., Ikeda, S., Koyano, S. et al. Identification of regulatory sites in the human PXR (NR1I2) promoter region. Mol Cell Biochem 281, 35–43 (2006). https://doi.org/10.1007/s11010-006-0167-7

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  • DOI: https://doi.org/10.1007/s11010-006-0167-7

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