Fission and uncoating of synaptic clathrin-coated vesicles are perturbed by disruption of interactions with the SH3 domain of endophilin.
Coordination between sequential steps in synaptic vesicle endocytosis, including clathrin coat formation, fission, and uncoating, appears to involve proteinprotein interactions. Here, we show that compounds that disrupt interactions of the SH3 domain of endophilin with dynamin and synaptojanin impair synaptic vesicle endocytosis in a living synapse. Two distinct endocytic intermediates accumulated. Free clathrin-coated vesicles were induced by a peptide-blocking endophilin's SH3 domain and by antibodies to the proline-rich domain (PRD) of synaptojanin. Invaginated clathrin-coated pits were induced by the same peptide and by the SH3 domain of endophilin. We suggest that the SH3 domain of endophilin participates in both fission and uncoating and that it may be a key component of a molecular switch that couples the fission reaction to uncoating.
Gad
H
H
The Nobel Institute for Neurophysiology, Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.
Ringstad
N
N
Löw
P
P
Kjaerulff
O
O
Gustafsson
J
J
Wenk
M
M
Di Paolo
G
G
Nemoto
Y
Y
Crun
J
J
Ellisman
M H
MH
De Camilli
P
P
Shupliakov
O
O
Brodin
L
L
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D002352:Carrier Proteins / Q000378:metabolism*; D002966:Clathrin / Q000378:metabolism*; D003001:Cloning, Molecular; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism*; D034281:Dynamins; D020558:GTP Phosphohydrolases / Q000378:metabolism; D007798:Lampreys; D008845:Microinjections; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010446:Peptide Fragments / Q000008:administration & dosage; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013572:Synaptic Vesicles / Q000378:metabolism*; D018909:src Homology Domains / Q000187:drug effects / Q000235:genetics / Q000502:physiology*
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D002966:Clathrin; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; C097702:synaptojanin; D010744:Phosphoric Monoester Hydrolases; D020558:GTP Phosphohydrolases; D034281:Dynamins