Galectin-3 drives glycosphingolipid-dependent biogenesis of clathrin-independent carriers.
Several cell surface molecules including signalling receptors are internalized by clathrin-independent endocytosis. How this process is initiated, how cargo proteins are sorted and membranes are bent remains unknown. Here, we found that a carbohydrate-binding protein, galectin-3 (Gal3), triggered the glycosphingolipid (GSL)-dependent biogenesis of a morphologically distinct class of endocytic structures, termed clathrin-independent carriers (CLICs). Super-resolution and reconstitution studies showed that Gal3 required GSLs for clustering and membrane bending. Gal3 interacted with a defined set of cargo proteins. Cellular uptake of the CLIC cargo CD44 was dependent on Gal3, GSLs and branched N-glycosylation. Endocytosis of β1-integrin was also reliant on Gal3. Analysis of different galectins revealed a distinct profile of cargoes and uptake structures, suggesting the existence of different CLIC populations. We conclude that Gal3 functionally integrates carbohydrate specificity on cargo proteins with the capacity of GSLs to drive clathrin-independent plasma membrane bending as a first step of CLIC biogenesis.
1] Institut Curie-Centre de Recherche, Endocytic Trafficking and Therapeutic Delivery group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [2] CNRS UMR3666, 75005 Paris, France [3] INSERM U1143, 75005 Paris, France [4] [5].
Ramya
R
Lakshminarayan
1] Institut Curie-Centre de Recherche, Endocytic Trafficking and Therapeutic Delivery group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [2] CNRS UMR3666, 75005 Paris, France [3] INSERM U1143, 75005 Paris, France [4] [5].
Christian
C
Wunder
1] Institut Curie-Centre de Recherche, Endocytic Trafficking and Therapeutic Delivery group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [2] CNRS UMR3666, 75005 Paris, France [3] INSERM U1143, 75005 Paris, France [4] [5].
Ulrike
U
Becken
Institute for Molecular Bioscience, University of Queensland, St Lucia, Queensland 4072, Australia.
Mark T
MT
Howes
Centre for Vascular Research, Australian Centre for Nanomedicine and ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, New South Wales 2052, Australia.
Carola
C
Benzing
1] Institut Curie-Centre de Recherche, Endocytic Trafficking and Therapeutic Delivery group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [2] CNRS UMR3666, 75005 Paris, France [3] INSERM U1143, 75005 Paris, France.
Senthil
S
Arumugam
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr. 108, 01307 Dresden, Germany.
Susanne
S
Sales
Institute for Molecular Bioscience, University of Queensland, St Lucia, Queensland 4072, Australia.
Nicholas
N
Ariotti
1] Institut Curie-Centre de Recherche, Endocytic Trafficking and Therapeutic Delivery group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [2] CNRS UMR3666, 75005 Paris, France [3] INSERM U1143, 75005 Paris, France [4].
Valérie
V
Chambon
1] CNRS UMR3666, 75005 Paris, France [2] INSERM U1143, 75005 Paris, France [3] Institut Curie-Centre de Recherche, Membrane Dynamics and Mechanics of Intracellular Signaling group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [4].
Christophe
C
Lamaze
Institut Curie-Centre de Recherche, Proteomics and Mass Spectrometry Laboratory, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
Damarys
D
Loew
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr. 108, 01307 Dresden, Germany.
Andrej
A
Shevchenko
Centre for Vascular Research, Australian Centre for Nanomedicine and ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, New South Wales 2052, Australia.
Katharina
K
Gaus
Institute for Molecular Bioscience, University of Queensland, St Lucia, Queensland 4072, Australia.
Robert G
RG
Parton
1] Institut Curie-Centre de Recherche, Endocytic Trafficking and Therapeutic Delivery group, 26 rue d'Ulm, 75248 Paris Cedex 05, France [2] CNRS UMR3666, 75005 Paris, France [3] INSERM U1143, 75005 Paris, France [4].
Ludger
L
Johannes
D000818:Animals; D001798:Blood Proteins; D004705:Endocytosis; D037502:Galectin 3 / Q000235:genetics / Q000378:metabolism*; D037161:Galectins; D006028:Glycosphingolipids / Q000378:metabolism*; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D051379:Mice; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D034622:RNA Interference; D014162:Transfection; D022161:Transport Vesicles / Q000378:metabolism*
D001798:Blood Proteins; C497483:CD44 protein, human; C476399:Cd44 protein, mouse; D037502:Galectin 3; D037161:Galectins; D006028:Glycosphingolipids; D018960:Hyaluronan Receptors; D019012:Integrin beta1; C428445:LGALS3 protein, human