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In\n                    <jats:italic>Drosophila<\/jats:italic>\n                    , Commissureless (Comm) plays a critical role in this process; however, there is conflicting data on the underlying molecular mechanism. Here, we demonstrate that the conserved PY motifs in the cytoplasmic domain of Comm are required allow the ubiquitination and lysosomal degradation of Robo1. Disruption of these motifs prevents Comm from localizing to Lamp1 positive late endosomes and to promote axon growth across the midline in vivo. In addition, we conclusively demonstrate a role for Nedd4 in midline crossing. Genetic analysis shows that\n                    <jats:italic>nedd4<\/jats:italic>\n                    mutations result in midline crossing defects in the\n                    <jats:italic>Drosophila<\/jats:italic>\n                    embryonic nerve cord, which can be rescued by introduction of exogenous Nedd4. Biochemical evidence shows that Nedd4 incorporates into a three-member complex with Comm and Robo1 in a PY motif-dependent manner. Finally, we present genetic evidence that Nedd4 acts with Comm in the embryonic nerve cord to downregulate Robo1 levels. Taken together, these findings demonstrate that Comm promotes midline crossing in the nerve cord by facilitating Robo1 ubiquitination by Nedd4, ultimately leading to its degradation.\n                  <\/jats:p>","DOI":"10.7554\/elife.92757","type":"journal-article","created":{"date-parts":[[2024,1,3]],"date-time":"2024-01-03T09:25:42Z","timestamp":1704273942000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.92757","source":"Crossref","is-referenced-by-count":1,"title":["Commissureless acts as a substrate adapter in a conserved Nedd4 E3 ubiquitin ligase pathway to promote axon growth across the midline"],"prefix":"10.7554","volume":"13","author":[{"given":"Kelly G","family":"Sullivan","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00b30xv10","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania","place":["Philadelphia, United States"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"conceptualization","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6146-0962","authenticated-orcid":true,"given":"Greg J","family":"Bashaw","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00b30xv10","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania","place":["Philadelphia, United States"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"corresponding-author","vocabulary":"crossref"},{"role":"conceptualization","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"resources","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"4374","published-online":{"date-parts":[[2025,5,23]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","first-page":"1019","DOI":"10.1002\/ajmg.b.30697","type":"journal-article","article-title":"Genetic analyses of roundabout (ROBO) axon guidance receptors in autism","volume":"147B","author":"Anitha","year":"2008","journal-title":"American Journal of Medical Genetics. 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