Robo recruitment of the Wave Regulatory Complex plays an essential and conserved role in midline repulsion
- University of Pennsylvania, United States
- Institut de recherches cliniques de Montréal (IRCM), Canada
Abstract
The Roundabout (Robo) guidance receptor family induces axon repulsion in response to its ligand Slit by inducing local cytoskeletal changes; however, the link to the cytoskeleton and the nature of these cytoskeletal changes are poorly understood. Here, we show that the heteropentameric Scar/Wave Regulatory Complex (WRC) which drives Arp2/3-induced branched actin polymerization, is a direct effector of Robo signaling. Biochemical evidence shows that Slit triggers WRC recruitment to the Robo receptor's WIRS motif. In Drosophila embryos, mutants of the WRC enhance Robo1-dependent midline crossing defects. Additionally, mutating Robo1's WIRS motif significantly reduces receptor activity in rescue assays in vivo, and CRISPR-Cas9 mutagenesis shows that the WIRS motif is essential for endogenous Robo1 function. Finally, axon guidance assays in mouse dorsal spinal commissural axons and gain-of-function experiments in chick embryos demonstrate that the WIRS motif is also required for Robo1 repulsion in mammals. Together, our data support an essential conserved role for the WIRS-WRC interaction in Robo1-mediated axon repulsion.
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All data generated or analysed during this study are included in the manuscript and supporting files.
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Author details
Funding
National Institutes of Health (R35 NS097340)
- Greg J Bashaw
National Science Foundation (IOS-1853719)
- Greg J Bashaw
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
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 of the Perelman School of Medicine at the University of Pennsylvania (Protocol #806216).
Copyright
© 2021, Chaudhari 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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Robo recruitment of the Wave Regulatory Complex plays an essential and conserved role in midline repulsion
eLife 10:e64474.
https://doi.org/10.7554/eLife.64474
