1. Structural Biology and Molecular Biophysics
Download icon

Estrogen receptor alpha somatic mutations Y537S and D538G confer breast cancer endocrine resistance by stabilizing the activating function-2 binding conformation

  1. Sean W Fanning
  2. Christopher G Mayne
  3. Venkatasubramanian Dharmarajan
  4. Kathryn E Carlson
  5. Teresa A Martin
  6. Scott J Novick
  7. Weiyi Toy
  8. Bradley Green
  9. Srinivas Panchamukhi
  10. Benita S Katzenellenbogen
  11. Emad Tajkhorshid
  12. Patrick R Griffin
  13. Yang Shen
  14. Sarat Chandarlapaty
  15. John A Katzenellenbogen
  16. Geoffrey L Griffin  Is a corresponding author
  1. University of Chicago, United States
  2. University of Illinois at Urbana-Champaign, United States
  3. The Scripps Research Institute-Scripps Florida, United States
  4. The Scripps Research Institute, United States
  5. Memorial Sloan Kettering Cancer Center, United States
  6. University of Illinois Urbana-Champaign, United States
  7. Texas A&M University, United States
  8. Memorial Sloan-Kettering Cancer Center, United States
Research Article
  • Cited 121
  • Views 5,789
  • Annotations
Cite this article as: eLife 2016;5:e12792 doi: 10.7554/eLife.12792

Abstract

Somatic mutations in the estrogen receptor alpha (ERα) gene (ESR1), especially Y537S and D538G, have been linked to acquired resistance to endocrine therapies. Cell based studies demonstrated that these mutants confer ERα constitutive activity and antiestrogen resistance and suggest that ligand-binding domain dysfunction leads to endocrine therapy resistance. Here, we integrate biophysical and structural biology data to reveal how these mutations lead to a constitutively active and antiestrogen resistant ERα. We show that these mutant ERs recruit coactivator in the absence of hormone while their affinities for estrogen agonist (estradiol) and antagonist (4-hydroxytamoxifen) are reduced. Further, they confer antiestrogen resistance by altering the conformational dynamics of the loop connecting Helix 11 and Helix 12 in the ligand-binding domain of ERα, which leads to a stabilized agonist state and an altered antagonist state that resists inhibition.

Article and author information

Author details

  1. Sean W Fanning

    Ben May Department for Cancer Research, University of Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Christopher G Mayne

    Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Venkatasubramanian Dharmarajan

    Department of Molecular Therapeutics, The Scripps Research Institute-Scripps Florida, Jupiter, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Kathryn E Carlson

    Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Teresa A Martin

    Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Scott J Novick

    Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Weiyi Toy

    Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Bradley Green

    Ben May Department for Cancer Research, University of Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Srinivas Panchamukhi

    Ben May Department for Cancer Research, University of Chicago, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Benita S Katzenellenbogen

    Department of Molecular and Integrative Physiology, University of Illinois Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Emad Tajkhorshid

    Department of Biochemistry, Center for Biophysics and Computational Biology, and Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Patrick R Griffin

    Department of Molecular Therapeutics, The Scripps Research Institute, Jupiter, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Yang Shen

    Department of Electrical and Computer Engineering and TEES-AgriLife Center for Bioinformatics and Genomic Systems Engineering, Texas A&M University, College Station, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Sarat Chandarlapaty

    Human Oncology and Pathogenesis Program, Memorial Sloan-Kettering Cancer Center, New York, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. John A Katzenellenbogen

    Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Geoffrey L Griffin

    Department of Molecular Therapeutics, University of Chicago, Jupiter, United States
    For correspondence
    ggreene@uchicago.edu
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Peter Tontonoz, Howard Hughes Medical Institute, University of California, Los Angeles, United States

Publication history

  1. Received: November 3, 2015
  2. Accepted: January 31, 2016
  3. Accepted Manuscript published: February 2, 2016 (version 1)
  4. Accepted Manuscript updated: February 3, 2016 (version 2)
  5. Version of Record published: March 24, 2016 (version 3)
  6. Version of Record updated: September 18, 2018 (version 4)

Copyright

© 2016, Fanning et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 5,789
    Page views
  • 1,392
    Downloads
  • 121
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, Scopus, PubMed Central.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

  1. Further reading

Further reading

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Quentin M Smith et al.
    Research Article

    Regulated thin filaments (RTFs) tightly control striated muscle contraction through calcium binding to troponin, which enables tropomyosin to expose myosin-binding sites on actin. Myosin binding holds tropomyosin in an open position, exposing more myosin-binding sites on actin, leading to cooperative activation. At lower calcium levels, troponin and tropomyosin turn off the thin filament; however, this is antagonised by the high local concentration of myosin, questioning how the thin filament relaxes. To provide molecular details of deactivation, we used single-molecule imaging of green fluorescent protein (GFP)-tagged myosin-S1 (S1-GFP) to follow the activation of RTF tightropes. In sub-maximal activation conditions, RTFs are not fully active, enabling direct observation of deactivation in real time. We observed that myosin binding occurs in a stochastic step-wise fashion; however, an unexpectedly large probability of multiple contemporaneous detachments is observed. This suggests that deactivation of the thin filament is a coordinated active process.

    1. Cell Biology
    2. Structural Biology and Molecular Biophysics
    Jesse R Holt et al.
    Research Article

    Keratinocytes, the predominant cell type of the epidermis, migrate to reinstate the epithelial barrier during wound healing. Mechanical cues are known to regulate keratinocyte re-epithelialization and wound healing however, the underlying molecular transducers and biophysical mechanisms remain elusive. Here, we show through molecular, cellular and organismal studies that the mechanically-activated ion channel PIEZO1 regulates keratinocyte migration and wound healing. Epidermal-specific Piezo1 knockout mice exhibited faster wound closure while gain-of-function mice displayed slower wound closure compared to littermate controls. By imaging the spatiotemporal localization dynamics of endogenous PIEZO1 channels we find that channel enrichment at some regions of the wound edge induces a localized cellular retraction that slows keratinocyte collective migration. In migrating single keratinocytes, PIEZO1 is enriched at the rear of the cell, where maximal retraction occurs, and we find that chemical activation of PIEZO1 enhances retraction during single as well as collective migration. Our findings uncover novel molecular mechanisms underlying single and collective keratinocyte migration that may suggest a potential pharmacological target for wound treatment. More broadly, we show that nanoscale spatiotemporal dynamics of Piezo1 channels can control tissue-scale events, a finding with implications beyond wound healing to processes as diverse as development, homeostasis, disease and repair.