Galectin-3 Mediated Endocytosis of the Orphan G-Protein-Coupled Receptor GPRC5A

Abstract Galectins, a family of glycan-binding proteins, play crucial roles in various cellular functions, acting at both intracellular and extracellular levels. Among them, Galectin-3 (Gal-3) stands out as a unique member, possessing an intrinsically disordered N-terminal interaction domain and a canonical carbohydrate-recognition domain (CRD). Gal-3 binding to glycosylated plasma membrane cargo leads to its oligomerization and membrane bending, ultimately resulting in the formation of endocytic invaginations. An interactomic assay using proteomic analysis of endogenous Gal-3 immunoprecipitates identified the orphan G protein-coupled receptor GPRC5A as a novel binding partner of Gal-3. GPRC5A, also known as Retinoic Acid-Induced protein 3 (RAI3), is transcriptionally induced by retinoic acid. Our results further demonstrate that extracellular recombinant Gal-3 stimulates GPRC5A internalization. In SW480 colorectal cancer cells, glycosylated GPRC5A interacts with Gal-3. Interestingly, while GPRC5A expression was upregulated by the addition of all-trans retinoic acid (ATRA), its endogenous internalization in SW480 cells was specifically triggered by extracellular Gal-3, but not by ATRA. This study provides new insights into the endocytic mechanisms of GPRC5A, for which no specific ligand has been identified to date. Further research may uncover additional Gal-3 mediated functions in GPRC5A cellular signaling and contribute to the development of innovative therapeutic strategies. Competing Interest Statement The authors have declared no competing interest. Footnotes boucheham{at}gmail.com (A.B.) ; j.mallor.f{at}gmail.com (J.M.F.) ; sbar{at}unistra.fr (S.B.) ; solene.zuttion{at}etu.unistra.fr (S.Z.) ; lana.blagec{at}gmx.de (L.B.) ; b.rinaldi{at}unistra.fr (B.R.) ; sbar{at}unistra.fr (S.B) ; solene.zuttion{at}etu.unistra.fr (S.Z.) ;, ewan.macdonald{at}crbm.cnrs.fr (E.M) ; Christian.Wunder{at}curie.fr (C.W.); ludger.johannes{at}curie.fr (L.J.),j.chicher{at}ibmc-cnrs.unistra.fr (J.C.);lauriane.kuhn{at}unistra.fr (L.K.); p.hammann{at}ibmc-cnrs.unistra.fr (P.H.)