The Phosphorylated Estrogen Receptor α (ER) Cistrome Identifies a Subset of Active Enhancers Enriched for Direct ER-DNA Binding and the Transcription Factor GRHL2

ABSTRACT

Posttranslational modifications are key regulators of protein function, providing cues that can alter protein interactions and cellular location. Phosphorylation of estrogen receptor α (ER) at serine 118 (pS118-ER) occurs in response to multiple stimuli and is involved in modulating ER-dependent gene transcription. While the cistrome of ER is well established, surprisingly little is understood about how phosphorylation impacts ER-DNA binding activity. To define the pS118-ER cistrome, chromatin immunoprecipitation sequencing was performed on pS118-ER and ER in MCF-7 cells treated with estrogen. pS118-ER occupied a subset of ER binding sites which were associated with an active enhancer mark, acetylated H3K27. Unlike ER, pS118-ER sites were enriched in GRHL2 DNA binding motifs, and estrogen treatment increased GRHL2 recruitment to sites occupied by pS118-ER. Additionally, pS118-ER occupancy sites showed greater enrichment of full-length estrogen response elements relative to ER sites. In an in vitro DNA binding array of genomic binding sites, pS118-ER was more commonly associated with direct DNA binding events than indirect binding events. These results indicate that phosphorylation of ER at serine 118 promotes direct DNA binding at active enhancers and is a distinguishing mark for associated transcription factor complexes on chromatin.

KEYWORDS:

• DNA arrays
• nuclear receptors
• protein phosphorylation
• protein-DNA interactions
• steroid receptors
• transcription factors

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Article of Significant Interest in This Issue

ACKNOWLEDGMENTS

We thank Amy Fowler for generously providing us with the MDA-MB-231 wt ER and S118A ER doxycycline-inducible cell lines. We also thank Audrey Gasch for helpful discussions regarding ChIP-seq analysis.

This work was supported by National Institutes of Health grant T32 CA009135 (to K.T.H.).

M.S.O. and C.L.W. are cofounders of Proteovista LLC.