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Research ArticleArticle

Dexamethasone Stimulates Human A1 Adenosine Receptor (A1AR) Gene Expression through Multiple Regulatory Sites in Promoter B

Hongzu Ren and Gary L. Stiles
Molecular Pharmacology February 1999, 55 (2) 309-316; DOI: https://doi.org/10.1124/mol.55.2.309
Hongzu Ren
Departments of Medicine and Pharmacology, Duke University Medical Center, Durham, North Carolina
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Gary L. Stiles
Departments of Medicine and Pharmacology, Duke University Medical Center, Durham, North Carolina
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Abstract

The expression of the human A1 adenosine receptor gene is controlled by two promoters, promoters A and B, and they are located 600 base pairs apart. The characteristics of the two promoters differ by the activity of expression, tissue specificity, and the potential regulatory elements around them. Promoter A is more active but its expression is observed only in selected tissues, whereas promoter B is constitutively expressed but at much reduced levels. In Chinese hamster ovary (CHO) cells transiently transfected with plasmids containing either promoter linked to a reporter gene, dexamethasone (dex) can stimulate (or enhance) the expression of promoter B much more effectively than that of promoter A. Mutation and deletion studies on plasmids containing promoter B have shown that the stimulation is mediated through multiple regulatory sites, including a serum response element, AP1, and TATA box. However, a single-glucocorticoid response element monomer-binding site between promoters A and B does not have significant contribution to dex-regulated expression. The interactions between glucocorticoid receptor (GR) and some regulatory sites are probably occurring via this protein (GR) interacting with other DNA-binding proteins because there is no GR DNA-binding sequence in the sites studied. The stimulation can be eliminated by mifepristone, an antagonist of GR, indicating the involvement of GR in gene regulation. In addition, dex treatment also stimulated the expression of A1 adenosine receptors in CHO cells transfected with the plasmids containing contiguous genomic sequences of promoter B or promoters A and B linked to the receptor-coding sequence. When promoter A is active and both promoter A and B are present in a construct, dex treatment induced a much smaller percentage of stimulation.

Footnotes

    • Received July 6, 1998.
    • Accepted November 10, 1998.
  • Send reprint requests to: Gary L. Stiles, M.D., Duke University Medical Center, Box 3444, Durham, NC 27710. E-mail:glsmd{at}duke.edu

  • This study was supported by National Heart, Lung and Blood Institute Specialized Center of Research Grant 5P50HL54314 in Ischemic Disease (G.L.S.).

  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 55 (2)
Molecular Pharmacology
Vol. 55, Issue 2
1 Feb 1999
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Research ArticleArticle

Dexamethasone Stimulates Human A1 Adenosine Receptor (A1AR) Gene Expression through Multiple Regulatory Sites in Promoter B

Hongzu Ren and Gary L. Stiles
Molecular Pharmacology February 1, 1999, 55 (2) 309-316; DOI: https://doi.org/10.1124/mol.55.2.309

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Research ArticleArticle

Dexamethasone Stimulates Human A1 Adenosine Receptor (A1AR) Gene Expression through Multiple Regulatory Sites in Promoter B

Hongzu Ren and Gary L. Stiles
Molecular Pharmacology February 1, 1999, 55 (2) 309-316; DOI: https://doi.org/10.1124/mol.55.2.309
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