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Transcriptional activities of estrogen and glucocorticoid receptors are functionally integrated at the AP-1 response
R M Uht1, C M Anderson, P Webb
1Department of Pathology, School of Medicine, University of California at San Francisco, 94143, USA.
Abstract:
Estrogens and glucocorticoids often act in opposition to regulate physiological responses. We investigated whether this might reflect the opposing actions of hormone-bound receptors on target genes regulated by the AP-1 response element. We performed a series of transfection experiments in which transcriptional activation, mediated by the AP-1 response element, was reflected in reporter gene activity. As previously described, we found that estrogens stimulate, whereas the glucocorticoid dexamethasone (Dex) inhibits, transcription through a model promoter from the collagenase gene (-73 to +63). This promoter bears a consensus AP-1 response element. When HeLa cells were treated with both estradiol and Dex, the steroids counteracted each other's transcriptional effects. The amount of transfected estrogen and glucocorticoid receptors (ER and GR) determined the extent to which Dex blunted estrogen stimulation or estrogen prevented Dex inhibition. The ER/GR interaction was observed both in the presence of estradiol and tamoxifen, which has previously been shown to have estrogen-like action at an AP-1 response element. The AP-1 family member c-Jun enhanced Dex inhibition and estradiol stimulation of transcriptional activation. c-Fos potentiated the effect of cotransfected c-Jun on estradiol stimulation but not Dex inhibition. The pattern of steroid responses was retained in the presence of the c-Jun activator phorbol 12-myristate 13-acetate. However, estradiol stimulation was lost in the presence of the c-Jun activator tumor necrosis factor-alpha. The ER/GR/AP-1 response element interaction was present, not only in a cell line originally derived from a uterine cervical adenocarcinoma (HeLa), but also in a cell line derived from the hypothalamus (GT1-1). Lastly, both progesterone receptor types A and B also interacted with the ER at the AP-1 site. These data indicate that opposing steroid influences can be mediated at the level of transcription through the AP-1 site and suggest that the integration of hormone action at this response element may underlie some of the opposing actions of estrogens and glucocorticoids or progestins on physiological responses.
Insights
Estrogen and glucocorticoid hormones oppose each other by interacting at the AP-1 response element, influencing gene transcription. This interaction, mediated by hormone receptors and AP-1 proteins, explains opposing physiological effects.
Area of Science:
- Molecular Endocrinology
- Gene Regulation
- Steroid Hormone Action
Background:
- Estrogens and glucocorticoids often exhibit opposing physiological effects.
- The molecular mechanisms underlying these opposing actions are not fully understood.
- Hormone receptors can influence gene transcription through specific DNA response elements.
Purpose of the Study:
- To investigate if opposing actions of estrogen and glucocorticoid receptors on target genes are mediated through the Activator Protein-1 (AP-1) response element.
- To elucidate the interaction between estrogen receptor (ER), glucocorticoid receptor (GR), and AP-1 proteins in regulating transcription.
Main Methods:
- Transfection experiments using reporter gene assays to measure transcriptional activation.
- Utilized a collagenase gene promoter with a consensus AP-1 response element.
- Investigated the effects of estradiol, dexamethasone (Dex), tamoxifen, AP-1 family members (c-Jun, c-Fos), and their activators on gene expression in HeLa and GT1-1 cells.
Main Results:
- Estrogens stimulated and dexamethasone inhibited transcription via the AP-1 site.
- ER and GR directly counteracted each other's effects on transcription.
- The extent of this antagonism was dependent on the levels of ER and GR.
- AP-1 proteins (c-Jun, c-Fos) modulated steroid receptor activity at the AP-1 site.
- Similar interactions were observed in both cervical adenocarcinoma (HeLa) and hypothalamic (GT1-1) cell lines.
- Progesterone receptors also interacted with ER at the AP-1 site.
Conclusions:
- Opposing steroid hormone actions can be mediated at the transcriptional level through the AP-1 response element.
- The integration of hormone receptor and AP-1 protein interactions at the AP-1 site likely underlies some opposing physiological effects of estrogens, glucocorticoids, and progestins.