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Research ArticleArticle

Transcriptional Regulation of Rat Hepatic Aryl Sulfotransferase (SULT1A1) Gene Expression by Glucocorticoids

Zhengbo Duanmu, Thomas A. Kocarek and Melissa Runge-Morris
Drug Metabolism and Disposition August 2001, 29 (8) 1130-1135;
Zhengbo Duanmu
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Thomas A. Kocarek
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Melissa Runge-Morris
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Abstract

The 5′-flanking region [1892 base pairs (bp)] of the rat aryl sulfotransferase (SULT1A1) gene was cloned and thecis-acting sequences involved in glucocorticoid-inducible SULT1A1 gene transcription were characterized. SULT1A1 promoter and 5′-flanking sequences lacked a TATA box and a consensus glucocorticoid response element. Using a 5′-rapid amplification of cDNA ends approach, four SULT1A1 transcription start sites were identified. Transient transfection studies with SULT1A1-5′:luciferase reporter constructs in primary cultured rat hepatocytes revealed that treatment with the potent glucocorticoid dexamethasone (10−9–10−5 M) produced concentration-dependent increases in luciferase activity in constructs containing from 1892 to 119 bp of the SULT1A1 5′-flanking region. Relative to the most upstream SULT1A1 transcription start site, the minimal cis-acting sequences that were required for dexamethasone-inducible SULT1A1 expression were located between −84 and −69 bp. Treatment of transfectants with a panel of steroids, including dexamethasone, triamcinolone acetonide, hydrocortisone, dihydrotestosterone, 17β-estradiol, and pregnenolone-16α-carbonitrile, revealed that steroid-inducible SULT1A1 gene expression was specific for glucocorticoid-class steroids. Concentration-response studies, coupled with a robust inhibition of glucocorticoid-inducible SULT1A1-5′:luciferase reporter activity by antiglucocorticoid/antiprogestin RU-486, recapitulated earlier findings on endogenous SULT1A1 gene expression and implicated a major role for the glucocorticoid receptor transcription factor in the regulation of glucocorticoid-inducible SULT1A1 gene expression.

Footnotes

  • This work was supported by National Institutes of Health Sciences Grants ES05823 (to M.R.M.) and HL50710 (to T.A.K.), and by services provided by the Cell Culture Facility Core and Imaging and Cytometry Facility Core of National Institute of Environmental Health Sciences Center Grant P30 ES06639.

  • Abbreviations used are::
    SULT1
    aryl sulfotransferase
    DEX
    dexamethasone
    PCR
    polymerase chain reaction
    GSP
    gene-specific primer
    RACE
    rapid amplification of cDNA ends
    bp
    base pair
    DMSO
    dimethyl sulfoxide
    GRE
    glucocorticoid response element
    ER3
    everted repeat with three intervening bases
    IR3
    inverted repeat with three intervening bases
    • Received February 20, 2001.
    • Accepted May 3, 2001.
  • The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 29 (8)
Drug Metabolism and Disposition
Vol. 29, Issue 8
1 Aug 2001
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Research ArticleArticle

Transcriptional Regulation of Rat Hepatic Aryl Sulfotransferase (SULT1A1) Gene Expression by Glucocorticoids

Zhengbo Duanmu, Thomas A. Kocarek and Melissa Runge-Morris
Drug Metabolism and Disposition August 1, 2001, 29 (8) 1130-1135;

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Research ArticleArticle

Transcriptional Regulation of Rat Hepatic Aryl Sulfotransferase (SULT1A1) Gene Expression by Glucocorticoids

Zhengbo Duanmu, Thomas A. Kocarek and Melissa Runge-Morris
Drug Metabolism and Disposition August 1, 2001, 29 (8) 1130-1135;
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