Ligand-induced large-scale chromatin dynamics as a biosensor for the detection of estrogen receptor subtype selective ligands

Rocio García-Becerra, Valeria Berno, David Ordaz-Rosado, Zelton D Sharp, Austin J. Cooney, Michael A. Mancini, Fernando Larrea

Research output: Contribution to journalArticle

4 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)37-44
Number of pages8
JournalGene
Volume458
Issue number1-2
DOIs
StatePublished - Jun 2010

Fingerprint

Biosensing Techniques
Estrogen Receptors
Chromatin
Ligands
HeLa Cells
Nandrolone
Steroids
Artificial Cells
Estrogen Receptor Modulators
Steroid Receptors
Progestins
Tamoxifen
Cytoplasmic and Nuclear Receptors
Fluorescent Antibody Technique
Estradiol
Estrogens
Transcription Factors
Genome

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 journalArticle

García-Becerra, R, Berno, V, Ordaz-Rosado, D, Sharp, ZD, Cooney, AJ, Mancini, MA & Larrea, F 2010, 'Ligand-induced large-scale chromatin dynamics as a biosensor for the detection of estrogen receptor subtype selective ligands', Gene, vol. 458, no. 1-2, pp. 37-44. https://doi.org/10.1016/j.gene.2010.03.007
García-Becerra, Rocio ; Berno, Valeria ; Ordaz-Rosado, David ; Sharp, Zelton D ; Cooney, Austin J. ; Mancini, Michael A. ; Larrea, Fernando. / Ligand-induced large-scale chromatin dynamics as a biosensor for the detection of estrogen receptor subtype selective ligands. In: Gene. 2010 ; Vol. 458, No. 1-2. pp. 37-44.
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