Structure and function of the ROR2 cysteine-rich domain in vertebrate noncanonical WNT5A signaling

  1. Samuel C Griffiths
  2. Jia Tan
  3. Armin Wagner
  4. Levi L Blazer
  5. Jarret J Adams
  6. Srisathya Srinivasan
  7. Shayan Moghisaei
  8. Sachdev S Sidhu
  9. Christian Siebold  Is a corresponding author
  10. Hsin-Yi Henry Ho  Is a corresponding author
  1. University of Oxford, United Kingdom
  2. University of California, Davis, United States
  3. Diamond Light Source, United Kingdom
  4. University of Waterloo, Canada

Abstract

The receptor tyrosine kinase ROR2 mediates noncanonical WNT5A signaling to orchestrate tissue morphogenetic processes, and dysfunction of the pathway causes Robinow syndrome, Brachydactyly B and metastatic diseases. The domain(s) and mechanisms required for ROR2 function, however, remain unclear. We solved the crystal structure of the extracellular cysteine-rich (CRD) and Kringle (Kr) domains of ROR2 and found that, unlike other CRDs, the ROR2 CRD lacks the signature hydrophobic pocket that binds lipids/lipid-modified proteins, such as WNTs, suggesting a novel mechanism of ligand reception. Functionally, we showed that the ROR2 CRD, but not other domains, is required and minimally sufficient to promote WNT5A signaling, and Robinow mutations in the CRD and the adjacent Kr impair ROR2 secretion and function. Moreover, using function-activating and -perturbing antibodies against the Frizzled (FZ) family of WNT receptors, we demonstrate the involvement of FZ in WNT5A-ROR signaling. Thus, ROR2 acts via its CRD to potentiate the function of a receptor super-complex that includes FZ to transduce WNT5A signals.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Samuel C Griffiths

    Division of Structural Biology, University of Oxford, Oxford, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  2. Jia Tan

    Department of Cell Biology and Human Anatomy, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Armin Wagner

    Science Division, Diamond Light Source, Didcot, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8995-7324
  4. Levi L Blazer

    School of Pharmacy, University of Waterloo, Waterloo, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9594-4642
  5. Jarret J Adams

    School of Pharmacy, University of Waterloo, Waterloo, Canada
    Competing interests
    The authors declare that no competing interests exist.
  6. Srisathya Srinivasan

    Department of Cell Biology and Human Anatomy, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Shayan Moghisaei

    Department of Cell Biology and Human Anatomy, University of California, Davis, Davis, United States
    Competing interests
    The authors declare that no competing interests exist.
  8. Sachdev S Sidhu

    School of Pharmacy, University of Waterloo, Waterloo, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7755-5918
  9. Christian Siebold

    Division of Structural Biology, University of Oxford, Oxford, United Kingdom
    For correspondence
    christian.siebold@strubi.ox.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
  10. Hsin-Yi Henry Ho

    Department of Cell Biology and Human Anatomy, University of California, Davis, Davis, United States
    For correspondence
    hyhho@ucdavis.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8780-7864

Funding

National Institutes of Health (1R35GM119574)

  • Hsin-Yi Henry Ho

National Institutes of Health (1R35GM144341)

  • Hsin-Yi Henry Ho

Cancer Research UK (C20724/A26752)

  • Christian Siebold

Wellcome Trust (203141/Z/16/Z)

  • Christian Siebold

Wellcome Trust (099675/Z/12/Z)

  • Samuel C Griffiths

National Institutes of Health (2P30CA093373-19)

  • Hsin-Yi Henry Ho

Cancer Research UK (DRCRPG-May23/100002)

  • Christian Siebold

National Institutes of Health (S10 OD018223)

  • Hsin-Yi Henry Ho

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

Reviewing Editor

  1. Olga Boudker, Weill Cornell Medicine, United States

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#21394) of the University of California, Davis. The protocol was approved by the IACUC of the University of California, Davis.

Version history

  1. Received: July 6, 2021
  2. Accepted: May 17, 2024
  3. Accepted Manuscript published: May 23, 2024 (version 1)

Copyright

© 2024, Griffiths 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. Samuel C Griffiths
  2. Jia Tan
  3. Armin Wagner
  4. Levi L Blazer
  5. Jarret J Adams
  6. Srisathya Srinivasan
  7. Shayan Moghisaei
  8. Sachdev S Sidhu
  9. Christian Siebold
  10. Hsin-Yi Henry Ho
(2024)
Structure and function of the ROR2 cysteine-rich domain in vertebrate noncanonical WNT5A signaling
eLife 13:e71980.
https://doi.org/10.7554/eLife.71980

Share this article

https://doi.org/10.7554/eLife.71980

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