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Analysis of glucocorticoid and androgen receptor gene fusions delineates domains required for transcriptional specificity

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Abstract

Androgen receptor (AR) and glucocorticoid receptor (GR) influence distinct physiologic responses in steroid-responsive cells despite their shared ability to selectively bind in vitro to the same canonical DNA sequence (TGTTCT). While the DNA-binding domains (DBDs) of these receptors are highly conserved, the amino N-terminal domain (NTD) and hormone-binding domain (HBD) are evolutionarily divergent. To determine the relative contribution of these functional domains to steroid-specific effects in vivo, we constructed a panel of AR/GR gene fusions by interchanging the NTD, DBD, and HBD regions of each receptor and measured transcriptional regulatory activities in transfected kidney and prostate cell lines. We found that GR was approximately 10-fold more active than AR when tested with the mouse mammary tumor virus promoter, and that this difference in activity was primarily owing to sequence divergence in the NTDs. We also tested transcriptional activation of the androgen-dependent rat probasin promoter, and in this case, AR was at least twofold more active than GR. Analysis of the chimeric receptors revealed that this difference mapped to the DBD region of the two receptors. Transcriptional repression functions of the wild-type and chimeric receptors were measured using an activator protein 1 (AP-1) transrepression assay and identified the GR HBD as a more potent transrepressor of AP-1 transcriptional activation than the AR HBD. Taken together, our analyses reveal that evolutionary sequence divergence between AR and GR functional domains results in unique promoter-specific activities within biologic systems in which both AR and GR are normally expressed.

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Authors and Affiliations

  1. Department of Molecular and Cellular Biology, University of Arizona, Tucson, AZ

    David C. Whitacre, K. Joy Karnas & Roger L. Miesfeld PhD

  2. Department of Biochemistry and Molecular Biophysics, Biological Sciences West 518A, University of Arizona, 85721, Tucson, AZ

    Roger L. Miesfeld PhD

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  1. David C. Whitacre
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  2. K. Joy Karnas
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Correspondence to Roger L. Miesfeld PhD.

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Whitacre, D.C., Karnas, K.J. & Miesfeld, R.L. Analysis of glucocorticoid and androgen receptor gene fusions delineates domains required for transcriptional specificity. Endocr 15, 111–118 (2001). https://doi.org/10.1385/ENDO:15:1:111

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  • DOI: https://doi.org/10.1385/ENDO:15:1:111

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