Interaction of 14-3-3 proteins with the Estrogen Receptor Alpha F domain provides a drug target interface

Ingrid J De Vries-van Leeuwen, Daniel da Costa Pereira, Koen D Flach, Sander R Piersma, Christian Haase, David Bier, Zeliha Yalcin, Rob Michalides, K Anton Feenstra, Connie R Jiménez, Tom F A de Greef, Luc Brunsveld, Christian Ottmann, Wilbert Zwart, Albertus H de Boer

Research output: Contribution to JournalArticle

Abstract

Estrogen receptor alpha (ERα) is involved in numerous physiological and pathological processes, including breast cancer. Breast cancer therapy is therefore currently directed at inhibiting the transcriptional potency of ERα, either by blocking estrogen production through aromatase inhibitors or antiestrogens that compete for hormone binding. Due to resistance, new treatment modalities are needed and as ERα dimerization is essential for its activity, interference with receptor dimerization offers a new opportunity to exploit in drug design. Here we describe a unique mechanism of how ERα dimerization is negatively controlled by interaction with 14-3-3 proteins at the extreme C terminus of the receptor. Moreover, the small-molecule fusicoccin (FC) stabilizes this ERα/14-3-3 interaction. Cocrystallization of the trimeric ERα/14-3-3/FC complex provides the structural basis for this stabilization and shows the importance of phosphorylation of the penultimate Threonine (ERα-T(594)) for high-affinity interaction. We confirm that T(594) is a distinct ERα phosphorylation site in the breast cancer cell line MCF-7 using a phospho-T(594)-specific antibody and by mass spectrometry. In line with its ERα/14-3-3 interaction stabilizing effect, fusicoccin reduces the estradiol-stimulated ERα dimerization, inhibits ERα/chromatin interactions and downstream gene expression, resulting in decreased cell proliferation. Herewith, a unique functional phosphosite and an alternative regulation mechanism of ERα are provided, together with a small molecule that selectively targets this ERα/14-3-3 interface.

Original languageEnglish
Pages (from-to)8894-9
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume110
Issue number22
DOIs
Publication statusPublished - 28 May 2013

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Keywords

  • 14-3-3 Proteins
  • Amino Acid Sequence
  • Breast Neoplasms
  • Crystallization
  • Dimerization
  • Drug Delivery Systems
  • Estrogen Receptor alpha
  • Female
  • Fluorescence Polarization
  • Gene Components
  • Gene Expression Regulation
  • Glycosides
  • Humans
  • MCF-7 Cells
  • Mass Spectrometry
  • Models, Molecular
  • Molecular Sequence Data
  • Phosphorylation
  • Protein Conformation
  • Protein Isoforms
  • Sequence Alignment
  • Journal Article
  • Research Support, Non-U.S. Gov't

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