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- Publisher
- Bioscientifica
- Copyright
- © 2009 Society for Endocrinology
- ISSN
- 0952-5041
- eISSN
- 1479-6813
- DOI
- 10.1677/JME-08-0177
- pmid
- 19376833
- Publisher site
- See Article on Publisher Site
Abstract
Estrogen receptors (ERs), ERα and ERβ, belong to a group of transcription factors that, upon ligand binding, regulate geneexpression by binding to specific DNA regions in chromatin as dimers. In this article, we applied the sequential chromatinimmunoprecipitation assay (Re-ChIP) to study the simultaneous presence of ERα and ERβ on various DNA-binding regions in intactchromatin. ERα/β heterodimers were isolated by precipitation with anti-ERβ antibody followed by anti-ERα antibody from a stableMCF-7-derived cell line that expresses endogenous ERα and an inducible version of ERβ. The Re-ChIP method was first validatedbased on the detection of ERα/β heterodimers bound to a promoter region of the pS2 gene known to bind both ERα and ERβ. We next examined 12 ER-binding sites using Re-ChIP assays for ERα/β heterodimer recruitment.Our results confirmed the recruitment of ERα/β heterodimers to all these regions. This study represents the first demonstrationof binding of ERα/β heterodimers to various DNA-binding regions in intact chromatin.
Journal
Journal of Molecular Endocrinology – Bioscientifica
Published: Aug 1, 2009