Abstract

Crosstalk between different receptor tyrosine kinases (RTKs) is thought to drive oncogenic signaling and allow therapeutic escape. EGFR and RON are two such RTKs from different subfamilies, which engage in crosstalk through unknown mechanisms. We combined high-resolution imaging with biochemical and mutational studies to ask how EGFR and RON communicate. EGF stimulation promotes EGFR-dependent phosphorylation of RON, but ligand stimulation of RON does not trigger EGFR phosphorylation – arguing that crosstalk is unidirectional. Nanoscale imaging reveals association of EGFR and RON in common plasma membrane microdomains. Two-color single particle tracking captured formation of complexes between RON and EGF-bound EGFR. Our results further show that RON is a substrate for EGFR kinase, and that transactivation of RON requires formation of a signaling competent EGFR dimer. These results support a role for direct EGFR/RON interactions in propagating crosstalk, such that EGF-stimulated EGFR phosphorylates RON to activate RON-directed signaling.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files. Source data for the quantitative plots and gels have been provided.

Article and author information

Author details

  1. Carolina Franco Nitta

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9122-5453
  2. Ellen W Green

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2083-522X
  3. Elton D Jhamba

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Justine M Keth

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Iraís Ortiz-Caraveo

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Rachel M Grattan

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. David J Schodt

    Department of Physics and Astronomy, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8986-2736
  8. Aubrey C Gibson

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Ashwani Rajput

    Department of Surgery, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Keith Lidke

    Department of Physics & Astronomy, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  11. Bridget S Wilson

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Mara P Steinkamp

    Department of Pathology, University of New Mexico, Albuquerque, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Diane S Lidke

    Department of Pathology, University of New Mexico, Albuquerque, United States
    For correspondence
    dlidke@salud.unm.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8533-6029

Funding

National Institutes of Health (R35GM126934)

  • Diane S Lidke

National Institutes of Health (R21GM132716)

  • Keith Lidke

National Institutes of Health (New Mexico Spatiotemporal Modeling Center,P50GM085273)

  • Bridget S Wilson
  • Diane S Lidke

National Institutes of Health (UNM MARC Program,2T34GM008751)

  • Justine M Keth

National Institutes of Health (ASERT-IRACDA,K12GM088021)

  • Elton D Jhamba

National Institutes of Health (UNM Undergraduate Pipeline Network,P30CA118100)

  • Justine M Keth
  • Aubrey C Gibson

National Institutes of Health (R01CA248166)

  • Diane S Lidke

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Satyajit Mayor, Marine Biological Laboratory, United States

Version history

  1. Preprint posted: August 12, 2020 (view preprint)
  2. Received: October 2, 2020
  3. Accepted: November 24, 2021
  4. Accepted Manuscript published: November 25, 2021 (version 1)
  5. Version of Record published: December 8, 2021 (version 2)

Copyright

© 2021, Franco Nitta 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.

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  1. Carolina Franco Nitta
  2. Ellen W Green
  3. Elton D Jhamba
  4. Justine M Keth
  5. Iraís Ortiz-Caraveo
  6. Rachel M Grattan
  7. David J Schodt
  8. Aubrey C Gibson
  9. Ashwani Rajput
  10. Keith Lidke
  11. Bridget S Wilson
  12. Mara P Steinkamp
  13. Diane S Lidke
(2021)
EGFR transactivates RON to drive oncogenic crosstalk
eLife 10:e63678.
https://doi.org/10.7554/eLife.63678

Share this article

https://doi.org/10.7554/eLife.63678

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