A fluorescent tagging approach in Drosophila reveals late endosomal trafficking of Notch and Sanpodo
Résumé
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.
Mots clés
APF
after puparium formation
BAC
bacterial artificial chromosome
Cad
cadherin
FRET
Forster resonance energy transfer
IQR
interquartile range
krz
kurtz
lgd
lethal(2) giant discs
Neur
Neural- ized
NICD
Notch intracellular domain
NiCherry
Notch intracellular Cherry
NiGFP
Notch intracellular GFP
Rbcn-3A
Rabconnectin-3A
sfGFP
superfolder GFP
SOP
sensory organ precursor cell
Spdo
Sanpodo
Domaines
Biologie du développement| Origine | Publication financée par une institution |
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