title: A fluorescent tagging approach in Drosophila reveals late endosomal trafficking of Notch and Sanpodo.
abstract: Signaling and endocytosis are highly integrated processes that regulate cell fate. In the Drosophila melanogaster sensory bristle lineages, Numb inhibits the recycling of Notch and its trafficking partner Sanpodo (Spdo) to regulate cell fate after asymmetric cell division. In this paper, we have used a dual GFP/Cherry tagging approach to study the distribution and endosomal sorting of Notch and Spdo in living pupae. The specific properties of GFP, i.e., quenching at low pH, and Cherry, i.e., slow maturation time, revealed distinct pools of Notch and Spdo: cargoes exhibiting high GFP/low Cherry fluorescence intensities localized mostly at the plasma membrane and early/sorting endosomes, whereas low GFP/high Cherry cargoes accumulated in late acidic endosomes. These properties were used to show that Spdo is sorted toward late endosomes in a Numb-dependent manner. This dual-tagging approach should be generally applicable to study the trafficking dynamics of membrane proteins in living cells and tissues.
authors.affiliation: Developmental and Stem Cell Biology Department, Institut Pasteur, 75015 Paris, France Centre National de la Recherche Scientifique, URA2578, 75015 Paris, France.
authors.forename: Lydie
authors.initials: L
authors.lastname: Couturier
authors.affiliation: Developmental and Stem Cell Biology Department, Institut Pasteur, 75015 Paris, France Centre National de la Recherche Scientifique, URA2578, 75015 Paris, France Master Biosciences, École Normale Supérieure de Lyon, 75015 Paris, France.
authors.forename: Mateusz
authors.initials: M
authors.lastname: Trylinski
authors.affiliation: Developmental and Stem Cell Biology Department, Institut Pasteur, 75015 Paris, France Centre National de la Recherche Scientifique, URA2578, 75015 Paris, France.
authors.forename: Khallil
authors.initials: K
authors.lastname: Mazouni
authors.affiliation: Developmental and Stem Cell Biology Department, Institut Pasteur, 75015 Paris, France Centre National de la Recherche Scientifique, URA2578, 75015 Paris, France.
authors.forename: Léa
authors.initials: L
authors.lastname: Darnet
authors.affiliation: Developmental and Stem Cell Biology Department, Institut Pasteur, 75015 Paris, France Centre National de la Recherche Scientifique, URA2578, 75015 Paris, France fschweis@pasteur.fr.
authors.forename: François
authors.initials: F
authors.lastname: Schweisguth
mesh_terms: D000818:Animals; D015820:Cadherins / Q000378:metabolism; D002455:Cell Division; D029721:Drosophila Proteins / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000166:cytology / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007605:Juvenile Hormones / Q000378:metabolism*; D008840:Microfilament Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D021381:Protein Transport; D051880:Receptors, Notch / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D059008:Time-Lapse Imaging
chemical_list: D015820:Cadherins; D029721:Drosophila Proteins; D007605:Juvenile Hormones; D008840:Microfilament Proteins; C497759:N protein, Drosophila; D051880:Receptors, Notch; D011993:Recombinant Fusion Proteins; C060606:numb protein, Drosophila; C112833:spdo protein, Drosophila; D049452:Green Fluorescent Proteins