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Submitted on March 29, 2005
Accepted on November 9, 2005
modulates ER
-mediated transcriptional activation by altering the recruitment of c-Fos and c-Jun to estrogen responsive promoters
Department of Biosciences at Novum, Karolinska Institutet, Novum, S-14157 Huddinge, Sweden; Department of Medical Nutrition, Karolinska Institutet, Novum, S-14186 Huddinge, Sweden
* To whom correspondence should be addressed. E-mail: jason.matthews{at}biosci.ki.se.
In this study, an ER
-expressing T47D cell line containing an inducible tet-off FLAG-ER
was used to examine the influence of ER on ER activity. Real-time PCR analysis of mRNA levels of two well studied estrogen responsive genes, pS2 and progesterone receptor (PR), showed that the expression levels of both genes were reduced in the presence of ER. Chromatin immunoprecipitation (ChIP) assays showed that the 17-estradiol (E2) induced recruitment patterns to the pS2 and PR promoters were similar for both ER and ER. ER expression did not significantly influence the kinetic recruitment profile of ER to the pS2 promoter, but it was evident that ER occupancy at the PR promoter was reduced. The E2-induced recruitment of c-Fos to a TRE site in the PR promoter was significantly reduced in the presence of ER, whereas only a slight reduction in the recruitment of c-Fos to the pS2 promoter was observed. ER expression resulted in a significant reduction in the E2 induced expression of c-Fos mRNA. The recruitment pattern of c-Jun was also altered by ER, although the expression levels of c-Jun were not. Expression of ER caused a further 30-50% decrease of the E2 induced reduction in ER protein after 3 h of E2 treatment, showing that ER influences ER protein levels. The altered recruitment of the AP-1 complex combined with the reduction in ER protein levels may partly explain the antagonistic effect of ER on ER-mediated transcription.
•
estrogen receptor •
activating protein-1 •
chromatin immunoprecipitation •
inhibition
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