Abstract
Estrogen receptors (ER), members of the nuclear steroid receptor superfamily, act to activate transcription through ligand-dependent recruitment of coregulators and chromatin modifications. A series of synthetic A-ring reduced 19-nortestosterone-derived progestins has the capacity to selectively bind ERΑ for activated transcription, and to recruit coregulatory factors. In this study, we have analyzed the ability of synthetic 19-nortestosterone derivatives to visibly alter the configuration of ER-target gene chromatin using a novel mammalian promoter transcriptional biosensor (PRL-array) stably transfected into the genome of HeLa cells (PRL-HeLa cells). Results from synthetic steroid-treated cells expressing functional GFP-ERΑ or YFP-ERΒ chimeras were compared to those obtained with estradiol (E2) and the antiestrogen tamoxifen. In the presence of synthetic ligands or E2 a concentration-dependent increase in area of the biosensor array was observed in GFP-ERΑ-expressing PRL-HeLa cells. No significant differences were found between the effects obtained with natural and synthetic steroids. Similarly, E2 or synthetic steroids-treated PRL-HeLa cells also resulted in similar colocalization of SRC-1- and RNAPII-immunofluorescence at the array. YFP-ERΒ-expressing PRL-HeLa cells treated with E2 showed increases in array area that were similar to ERΑ however, treatment of YFP-ERΒ-expressing cells with synthetic ligands was indistinguishable from vehicle controls. These data indicate that A-ring reduced 19-nortestosterone derivatives have an estrogen-like effect on chromatin, including recruitment of transcription factors through selective interactions with ERΑ.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 37-44 |
| Number of pages | 8 |
| Journal | Gene |
| Volume | 458 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Jun 2010 |
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Keywords
- Chromatin
- Estrogen receptor subtypes
- Ligand selectivity
- Synthetic progestins
- Transcription
ASJC Scopus subject areas
- Genetics
Cite this
Ligand-induced large-scale chromatin dynamics as a biosensor for the detection of estrogen receptor subtype selective ligands. / García-Becerra, Rocio; Berno, Valeria; Ordaz-Rosado, David; Sharp, Zelton D; Cooney, Austin J.; Mancini, Michael A.; Larrea, Fernando.
In: Gene, Vol. 458, No. 1-2, 06.2010, p. 37-44.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Ligand-induced large-scale chromatin dynamics as a biosensor for the detection of estrogen receptor subtype selective ligands
AU - García-Becerra, Rocio
AU - Berno, Valeria
AU - Ordaz-Rosado, David
AU - Sharp, Zelton D
AU - Cooney, Austin J.
AU - Mancini, Michael A.
AU - Larrea, Fernando
PY - 2010/6
Y1 - 2010/6
N2 - Estrogen receptors (ER), members of the nuclear steroid receptor superfamily, act to activate transcription through ligand-dependent recruitment of coregulators and chromatin modifications. A series of synthetic A-ring reduced 19-nortestosterone-derived progestins has the capacity to selectively bind ERΑ for activated transcription, and to recruit coregulatory factors. In this study, we have analyzed the ability of synthetic 19-nortestosterone derivatives to visibly alter the configuration of ER-target gene chromatin using a novel mammalian promoter transcriptional biosensor (PRL-array) stably transfected into the genome of HeLa cells (PRL-HeLa cells). Results from synthetic steroid-treated cells expressing functional GFP-ERΑ or YFP-ERΒ chimeras were compared to those obtained with estradiol (E2) and the antiestrogen tamoxifen. In the presence of synthetic ligands or E2 a concentration-dependent increase in area of the biosensor array was observed in GFP-ERΑ-expressing PRL-HeLa cells. No significant differences were found between the effects obtained with natural and synthetic steroids. Similarly, E2 or synthetic steroids-treated PRL-HeLa cells also resulted in similar colocalization of SRC-1- and RNAPII-immunofluorescence at the array. YFP-ERΒ-expressing PRL-HeLa cells treated with E2 showed increases in array area that were similar to ERΑ however, treatment of YFP-ERΒ-expressing cells with synthetic ligands was indistinguishable from vehicle controls. These data indicate that A-ring reduced 19-nortestosterone derivatives have an estrogen-like effect on chromatin, including recruitment of transcription factors through selective interactions with ERΑ.
AB - Estrogen receptors (ER), members of the nuclear steroid receptor superfamily, act to activate transcription through ligand-dependent recruitment of coregulators and chromatin modifications. A series of synthetic A-ring reduced 19-nortestosterone-derived progestins has the capacity to selectively bind ERΑ for activated transcription, and to recruit coregulatory factors. In this study, we have analyzed the ability of synthetic 19-nortestosterone derivatives to visibly alter the configuration of ER-target gene chromatin using a novel mammalian promoter transcriptional biosensor (PRL-array) stably transfected into the genome of HeLa cells (PRL-HeLa cells). Results from synthetic steroid-treated cells expressing functional GFP-ERΑ or YFP-ERΒ chimeras were compared to those obtained with estradiol (E2) and the antiestrogen tamoxifen. In the presence of synthetic ligands or E2 a concentration-dependent increase in area of the biosensor array was observed in GFP-ERΑ-expressing PRL-HeLa cells. No significant differences were found between the effects obtained with natural and synthetic steroids. Similarly, E2 or synthetic steroids-treated PRL-HeLa cells also resulted in similar colocalization of SRC-1- and RNAPII-immunofluorescence at the array. YFP-ERΒ-expressing PRL-HeLa cells treated with E2 showed increases in array area that were similar to ERΑ however, treatment of YFP-ERΒ-expressing cells with synthetic ligands was indistinguishable from vehicle controls. These data indicate that A-ring reduced 19-nortestosterone derivatives have an estrogen-like effect on chromatin, including recruitment of transcription factors through selective interactions with ERΑ.
KW - Chromatin
KW - Estrogen receptor subtypes
KW - Ligand selectivity
KW - Synthetic progestins
KW - Transcription
UR - http://www.scopus.com/inward/record.url?scp=77952550322&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77952550322&partnerID=8YFLogxK
U2 - 10.1016/j.gene.2010.03.007
DO - 10.1016/j.gene.2010.03.007
M3 - Article
C2 - 20347019
AN - SCOPUS:77952550322
VL - 458
SP - 37
EP - 44
JO - Gene
JF - Gene
SN - 0378-1119
IS - 1-2
ER -