1. Developmental Biology
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An RNAi screen unravels the complexities of Rho GTPase networks in skin morphogenesis

  1. Melanie Laurin
  2. Nicholas C Gomez
  3. John Levorse
  4. Ataman Sendoel
  5. Megan Sribour
  6. Elaine Fuchs  Is a corresponding author
  1. The Rockefeller University, United States
Research Article
  • Cited 4
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Cite this article as: eLife 2019;8:e50226 doi: 10.7554/eLife.50226

Abstract

During mammalian embryogenesis, extensive cellular remodeling is needed for tissue morphogenesis. As effectors of cytoskeletal dynamics, Rho GTPases and their regulators are likely involved, but their daunting complexity has hindered progress in dissecting their functions. We overcome this hurdle by employing high throughput in utero RNAi-mediated screening to identify key Rho regulators of skin morphogenesis. Our screen unveiled hitherto unrecognized roles for Rho-mediated cytoskeletal remodeling events that impact hair follicle specification, differentiation, downgrowth and planar cell polarity. Coupling our top hit with gain/loss-of-function genetics, interactome proteomics and tissue imaging, we show that RHOU, an atypical Rho, governs the cytoskeletal-junction dynamics that establish columnar shape and planar cell polarity in epidermal progenitors. Conversely, RHOU downregulation is required to remodel to a conical cellular shape that enables hair bud invagination and downgrowth. Our findings underscore the power of coupling screens with proteomics to unravel the physiological significance of complex gene families.

Data availability

Sequencing data have been deposited in NCBI GEO under accession number GSE123047. All data generated or analysed during this study are included in the manuscript and supporting files

The following data sets were generated

Article and author information

Author details

  1. Melanie Laurin

    Robin Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  2. Nicholas C Gomez

    Robin Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  3. John Levorse

    Robin Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  4. Ataman Sendoel

    Robin Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  5. Megan Sribour

    Robin Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States
    Competing interests
    No competing interests declared.
  6. Elaine Fuchs

    Robin Neustein Laboratory of Mammalian Cell Biology and Development, The Rockefeller University, New York, United States
    For correspondence
    fuchs@rockefeller.edu
    Competing interests
    Elaine Fuchs, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7198-3257

Funding

Howard Hughes Medical Institute

  • Elaine Fuchs

Canadian Institutes of Health Research

  • Melanie Laurin

Burroughs Wellcome Fund (Postdoctoral Enrichment Program Award)

  • Nicholas C Gomez

National Institutes of Health (Ruth L. Kirschstein National Research Service Award F32CA221353)

  • Nicholas C Gomez

Human Frontier Science Program

  • Ataman Sendoel

Marie Curie Foundation

  • Ataman Sendoel

Cancer Research Society

  • Melanie Laurin

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 mouse strains were housed in an AAALAC-accredited facility and experiments were conducted according to the Rockefeller University's Institutional Animal Care and Use Committee, and NIH guidelines for Animal Care and Use.All animal procedures used in this study are described in our #17020-H protocol named Lentiviral RNAi and Skin, which had been previously reviewed and approved by the Rockefeller University Institutional Animal Care and Use Committee (IACUC)

Reviewing Editor

  1. Maddy Parsons, King's College London, United Kingdom

Publication history

  1. Received: July 16, 2019
  2. Accepted: September 24, 2019
  3. Accepted Manuscript published: September 25, 2019 (version 1)
  4. Version of Record published: September 30, 2019 (version 2)

Copyright

© 2019, Laurin 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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