APC regulates NSLIT2-induced internalization and signaling of the chemorepellent receptor, ROBO1
The SLIT–Roundabout signaling pathway regulates axon guidance and cell migration, and ROBO1 is a receptor for SLIT ligands. ROBO1 undergoes constitutive endocytosis, which is enhanced upon NSLIT2 binding, but the molecular mechanisms and functional consequences of this process are not well understood. Using pharmacologic inhibitors and molecular techniques, we found that clathrin-mediated endocytosis is necessary for NSLIT2-induced inhibition of cell spreading. To explore the underlying mechanisms, we performed proximity-dependent biotin identification to identify ROBO1-interacting proteins whose association with the cytoplasmic domain of ROBO1 is differentially regulated by NSLIT2. We discovered that adenomatous polyposis coli, a multifunctional tumor suppressor, is in close proximity to ROBO1 but dissociates upon binding of NSLIT2 and that this dissociation is necessary for clathrin-mediated endocytosis of ROBO1 and subsequent effects on cell morphology. These findings provide new insights into the functional mechanisms by which NSLIT2 binding to ROBO1 affects changes in actin cytoskeletal architecture.
Authors
- Brian Raught (ORCID: https://orcid.org/0000-0001-6145-4776)
- Étienne Coyaud (ORCID: https://orcid.org/0000-0002-5893-4557)
- Lisa Annette Robinson (ORCID: https://orcid.org/0000-0003-1714-1929)
- Sergio Grinstein (ORCID: https://orcid.org/0000-0002-0795-4160)
- Yiwei Huang (ORCID: https://orcid.org/0000-0001-7294-878X)
- Richard F. Collins (ORCID: https://orcid.org/0000-0002-0670-7424)
- Zhubing Li (ORCID: https://orcid.org/0000-0002-2432-2245)
- Sajedabanu Patel (ORCID: https://orcid.org/0000-0003-3798-3306)
- Amir Mohamed (ORCID: https://orcid.org/0000-0002-1141-1935)
- Nourhan R. Almasri (ORCID: https://orcid.org/0009-0001-8036-3439)
- Wenjuan Li
- Jonathan St-Germain
- Bo Wen Pang
Institutions
- University Health Network (CA)
- Inserm (FR)
- University of Toronto (CA)
- Université de Lille (FR)
- Great Ormond Street Hospital (GB)
- Hospital for Sick Children (CA)
- Princess Margaret Cancer Centre (CA)
- Protéomique, Réponse Inflammatoire et Spectrométrie de Masse (FR)
Publication Details
- Journal
- Life Science Alliance
- Published
- 2026-11-11
- DOI
- https://doi.org/10.26508/lsa.202403190
- Citations
- 1
- Primary Topic
- Axon Guidance and Neuronal Signaling
- Type
- article
- Field-Weighted Citation Impact
- 1.43
Funders
- Hospital for Sick Children
- Canadian Institutes of Health Research
- Natural Sciences and Engineering Research Council of Canada