Recruitment of activated G protein-coupled receptors to pre-existing clathrin-coated pits in living cells.
The process of clathrin-mediated endocytosis tightly regulates signaling of the superfamily of seven-transmembrane G protein-coupled receptors (GPCRs). A fundamental question in the cell biology of membrane receptor endocytosis is whether activated receptors can initiate the formation of clathrin-coated pits (CPs) or whether they are simply mobilized to pre-existing CPs. Here, using various approaches, including a dynamic assay to monitor the distribution of CPs and GPCR-beta-arrestin complexes in live HeLa cells, we demonstrate for the first time that activated GPCRs do not initiate the de novo formation of CPs but instead are targeted to pre-existing CPs.
Scott
Mark G H
MG
Department of Cell Biology, Institut Cochin de Génétique Moléculaire, Pavillon Gustave Roussy, 75679 Paris CEDEX 14, France.
Benmerah
Alexandre
A
Muntaner
Olivier
O
Marullo
Stefano
S
D019390:Arrestins / Q000235:genetics / Q000502:physiology; D001426:Bacterial Proteins / Q000235:genetics; D022163:Clathrin-Coated Vesicles / Q000502:physiology* / Q000648:ultrastructure; D019204:GTP-Binding Proteins / Q000378:metabolism*; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics; D010957:Plasmids; D011956:Receptors, Cell Surface / Q000502:physiology*; D018025:Receptors, Thyrotropin-Releasing Hormone / Q000502:physiology; D011990:Receptors, Transferrin / Q000235:genetics / Q000502:physiology; D011994:Recombinant Proteins / Q000378:metabolism; D013347:Subcellular Fractions / Q000502:physiology; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology; D014162:Transfection; D000071557:beta-Arrestins
D019390:Arrestins; D001426:Bacterial Proteins; D008164:Luminescent Proteins; D011956:Receptors, Cell Surface; D018025:Receptors, Thyrotropin-Releasing Hormone; D011990:Receptors, Transferrin; D011994:Recombinant Proteins; D000071557:beta-Arrestins; C054562:yellow fluorescent protein, Bacteria; D013973:Thyrotropin-Releasing Hormone; D019204:GTP-Binding Proteins