title: Vesicle uncoating regulated by SH3-SH3 domain-mediated complex formation between endophilin and intersectin at synapses.
abstract: Neurotransmission involves the exo-endocytic cycling of synaptic vesicle (SV) membranes. Endocytic membrane retrieval and clathrin-mediated SV reformation require curvature-sensing and membrane-bending BAR domain proteins such as endophilin A. While their ability to sense and stabilize curved membranes facilitates membrane recruitment of BAR domain proteins, the precise mechanisms by which they are targeted to specific sites of SV recycling has remained unclear. Here, we demonstrate that the multi-domain scaffold intersectin 1 directly associates with endophilin A to facilitate vesicle uncoating at synapses. Knockout mice deficient in intersectin 1 accumulate clathrin-coated vesicles at synapses, a phenotype akin to loss of endophilin function. Intersectin 1/endophilin A1 complex formation is mediated by direct binding of the SH3B domain of intersectin to a non-canonical site on the SH3 domain of endophilin A1. Consistent with this, intersectin-binding defective mutant endophilin A1 fails to rescue clathrin accumulation at neuronal synapses derived from endophilin A1-3 triple knockout (TKO) mice. Our data support a model in which intersectin aids endophilin A recruitment to sites of clathrin-mediated SV recycling, thereby facilitating vesicle uncoating.
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany Department of Neuroscience, DBRM, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Arndt
authors.initials: A
authors.lastname: Pechstein
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany.
authors.forename: Fabian
authors.initials: F
authors.lastname: Gerth
authors.affiliation: European Neuroscience Institute Göttingen (ENI-G), Synaptic Vesicle Dynamics Group, Göttingen, Germany.
authors.forename: Ira
authors.initials: I
authors.lastname: Milosevic
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Jäpel
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.
authors.forename: Marielle
authors.initials: M
authors.lastname: Eichhorn-Grünig
authors.affiliation: Department of Neuroscience, DBRM, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Olga
authors.initials: O
authors.lastname: Vorontsova
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.
authors.forename: Jelena
authors.initials: J
authors.lastname: Bacetic
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Maritzen
authors.affiliation: Department of Neuroscience, DBRM, Karolinska Institutet, Stockholm, Sweden oleg.shupliakov@ki.se christian.freund@fu-berlin.de haucke@fmp-berlin.de.
authors.forename: Oleg
authors.initials: O
authors.lastname: Shupliakov
authors.affiliation: Institute of Chemistry and Biochemistry, Freie Universität Berlin, Berlin, Germany oleg.shupliakov@ki.se christian.freund@fu-berlin.de haucke@fmp-berlin.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Freund
authors.affiliation: Leibniz-Institut für Molekulare Pharmakologie (FMP), Berlin, Germany oleg.shupliakov@ki.se christian.freund@fu-berlin.de haucke@fmp-berlin.de.
authors.forename: Volker
authors.initials: V
authors.lastname: Haucke
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism; D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D022163:Clathrin-Coated Vesicles / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D018345:Mice, Knockout; D018613:Microscopy, Confocal; D013569:Synapses / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; C504641:endophilin A1 protein, mouse; C115919:intersectin 1; D000217:Acyltransferases; C018764:2-acylglycerophosphate acyltransferase
keywords: SH3 domains; endophilin; intersectin; neurotransmission; synaptic vesicle recycling