1. Developmental Biology
  2. Neuroscience

Plexins promote hedgehog signaling through their cytoplasmic GAP activity

  1. Justine M Pinskey
  2. Tyler M Hoard
  3. Xiao-Feng Zhao
  4. Nicole E Franks
  5. Zoë C Frank
  6. Alexandra N McMellen
  7. Roman J Giger
  8. Benjamin L Allen  Is a corresponding author
  1. University of Michigan-Ann Arbor, United States
https://doi.org/10.7554/eLife.74750
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Cite this article
  1. Justine M Pinskey
  2. Tyler M Hoard
  3. Xiao-Feng Zhao
  4. Nicole E Franks
  5. Zoë C Frank
  6. Alexandra N McMellen
  7. Roman J Giger
  8. Benjamin L Allen
(2022)
Plexins promote hedgehog signaling through their cytoplasmic GAP activity
eLife 11:e74750.
https://doi.org/10.7554/eLife.74750
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Abstract

Hedgehog signaling controls tissue patterning during embryonic and postnatal development and continues to play important roles throughout life. Characterizing the full complement of Hedgehog pathway components is essential to understanding its wide-ranging functions. Previous work has identified Neuropilins, established Semaphorin receptors, as positive regulators of Hedgehog signaling. Neuropilins require Plexin co-receptors to mediate Semaphorin signaling, but a role for Plexins in Hedgehog signaling has not yet been explored. Here, we provide evidence that multiple Plexins promote Hedgehog signaling in NIH/3T3 mouse fibroblasts and that Plexin loss-of-function in these cells results in significantly reduced Hedgehog pathway activity. Catalytic activity of the Plexin GTPase activating protein (GAP) domain is required for Hedgehog signal promotion, and constitutive activation of the GAP domain further amplifies Hedgehog signaling. Additionally, we demonstrate that Plexins promote Hedgehog signaling at the level of GLI transcription factors and that this promotion requires intact primary cilia. Finally, we find that Plexin loss-of-function significantly reduces the response to Hedgehog pathway activation in the mouse dentate gyrus. Together, these data identify Plexins as novel components of the Hedgehog pathway and provide insight into their mechanism of action.

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

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Author details

  1. Justine M Pinskey

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Tyler M Hoard

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Xiao-Feng Zhao

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7574-7163

  4. Nicole E Franks

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Zoë C Frank

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Alexandra N McMellen

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Roman J Giger

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2926-3336

  8. Benjamin L Allen

    Department of Cell and Developmental Biology, University of Michigan-Ann Arbor, Ann Arbor, United States
    For correspondence
    benallen@umich.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2323-8313

Funding

National Institutes of Health (R01DC014428)

  • Benjamin L Allen

National Institutes of Health (R01CA198074)

  • Benjamin L Allen

National Institutes of Health (R01GM118751)

  • Benjamin L Allen

National Institutes of Health (R01MH119346)

  • Roman J Giger

National Institutes of Health (F31NS096734)

  • Justine M Pinskey

National Institutes of Health (T32HD007505)

  • Justine M Pinskey

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

Ethics

Animal experimentation: All mice were housed in specific pathogen-free facilities at the University of Michigan. This study was approved by the University of Michigan Institutional Animal Care and Use Committee (IACUC; Protocol Number: PRO00010440).

Reviewing Editor

  1. Jeremy F Reiter, University of California, San Francisco, United States

Publication history

  1. Received: October 15, 2021
  2. Preprint posted: December 16, 2021 (view preprint)
  3. Accepted: September 27, 2022
  4. Accepted Manuscript published: September 28, 2022 (version 1)
  5. Version of Record published: October 11, 2022 (version 2)

Copyright

© 2022, Pinskey 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. Justine M Pinskey
  2. Tyler M Hoard
  3. Xiao-Feng Zhao
  4. Nicole E Franks
  5. Zoë C Frank
  6. Alexandra N McMellen
  7. Roman J Giger
  8. Benjamin L Allen
(2022)
Plexins promote hedgehog signaling through their cytoplasmic GAP activity
eLife 11:e74750.
https://doi.org/10.7554/eLife.74750

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