Abstract
Clathrin-mediated endocytosis, the major pathway for ligand internalization into eukaryotic cells, is thought to be initiated by the clustering of clathrin and adaptors around receptors destined for internalization. However, here we report that the membrane-sculpting F-BAR domain-containing Fer/Cip4 homology domain-only proteins 1 and 2 (FCHo1/2) were required for plasma membrane clathrin-coated vesicle (CCV) budding and marked sites of CCV formation. Changes in FCHo1/2 expression levels correlated directly with numbers of CCV budding events, ligand endocytosis, and synaptic vesicle marker recycling. FCHo1/2 proteins bound specifically to the plasma membrane and recruited the scaffold proteins eps15 and intersectin, which in turn engaged the adaptor complex AP2. The FCHo F-BAR membrane-bending activity was required, leading to the proposal that FCHo1/2 sculpt the initial bud site and recruit the clathrin machinery for CCV formation.
MeSH Terms
Adaptor Protein Complex 2/metabolism
Adaptor Proteins, Signal Transducing
Adaptor Proteins, Vesicular Transport/metabolism
Animals
Calcium-Binding Proteins/metabolism
Cell Line
Cell Membrane/metabolism
Cells, Cultured
Clathrin/metabolism
Clathrin-Coated Vesicles/metabolism
Endocytosis
Fatty Acid-Binding Proteins
HeLa Cells
Humans
Intracellular Signaling Peptides and Proteins/metabolism
Membrane Proteins
Mice
Models, Molecular
Neurons/cytology,metabolism
Phosphoproteins/metabolism
Protein Multimerization
Protein Structure, Tertiary
Proteins/chemistry,metabolism
RNA Interference
Rats
Rats, Sprague-Dawley
Recombinant Fusion Proteins/metabolism
Synaptic Vesicles/metabolism
Chemicals
Adaptor Protein Complex 2
Adaptor Proteins, Signal Transducing
Adaptor Proteins, Vesicular Transport
Calcium-Binding Proteins
Clathrin
EPS15 protein, human
FCHO1 protein, human
Fatty Acid-Binding Proteins
Fcho2 protein, mouse
Intracellular Signaling Peptides and Proteins
Membrane Proteins
Phosphoproteins
Proteins
Recombinant Fusion Proteins
intersectin 1
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Henne William Mike
Medical Research Council, Laboratory of Molecular Biology (MRC-LMB), Hills Road, Cambridge CB2 0QH, UK.
Boucrot Emmanuel
Meinecke Michael
Evergren Emma
Vallis Yvonne
Mittal Rohit
McMahon Harvey T
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