Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion
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 Reg...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2021-04-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/64474 |
| _version_ | 1818028344561631232 |
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| author | Karina Chaudhari Madhavi Gorla Chao Chang Artur Kania Greg J Bashaw |
| author_facet | Karina Chaudhari Madhavi Gorla Chao Chang Artur Kania Greg J Bashaw |
| author_sort | Karina Chaudhari |
| collection | DOAJ |
| description | 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 WRC-interacting receptor sequence (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. |
| first_indexed | 2024-12-10T05:02:18Z |
| format | Article |
| id | doaj.art-c69636800f26467780990d10d50246b7 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-12-10T05:02:18Z |
| publishDate | 2021-04-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-c69636800f26467780990d10d50246b72022-12-22T02:01:21ZengeLife Sciences Publications LtdeLife2050-084X2021-04-011010.7554/eLife.64474Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsionKarina Chaudhari0https://orcid.org/0000-0003-3533-3027Madhavi Gorla1Chao Chang2Artur Kania3https://orcid.org/0000-0002-5209-2520Greg J Bashaw4https://orcid.org/0000-0002-6146-0962Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania, Philadelphia, United StatesDepartment of Neuroscience, Perelman School of Medicine, University of Pennsylvania, Philadelphia, United StatesInstitut de recherches cliniques de Montréal (IRCM), Montréal, Canada; Department of Anatomy and Cell Biology and Division of Experimental Medicine, McGill University, Montréal, CanadaInstitut de recherches cliniques de Montréal (IRCM), Montréal, Canada; Department of Anatomy and Cell Biology and Division of Experimental Medicine, McGill University, Montréal, CanadaDepartment of Neuroscience, Perelman School of Medicine, University of Pennsylvania, Philadelphia, United StatesThe 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 WRC-interacting receptor sequence (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.https://elifesciences.org/articles/64474axon guidancespinal cordmidlineSlitRobowave regulatory complex |
| spellingShingle | Karina Chaudhari Madhavi Gorla Chao Chang Artur Kania Greg J Bashaw Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion eLife axon guidance spinal cord midline Slit Robo wave regulatory complex |
| title | Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion |
| title_full | Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion |
| title_fullStr | Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion |
| title_full_unstemmed | Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion |
| title_short | Robo recruitment of the Wave regulatory complex plays an essential and conserved role in midline repulsion |
| title_sort | robo recruitment of the wave regulatory complex plays an essential and conserved role in midline repulsion |
| topic | axon guidance spinal cord midline Slit Robo wave regulatory complex |
| url | https://elifesciences.org/articles/64474 |
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