title: Recruitment of endophilin to clathrin-coated pit necks is required for efficient vesicle uncoating after fission.
abstract: Endophilin is a membrane-binding protein with curvature-generating and -sensing properties that participates in clathrin-dependent endocytosis of synaptic vesicle membranes. Endophilin also binds the GTPase dynamin and the phosphoinositide phosphatase synaptojanin and is thought to coordinate constriction of coated pits with membrane fission (via dynamin) and subsequent uncoating (via synaptojanin). We show that although synaptojanin is recruited by endophilin at bud necks before fission, the knockout of all three mouse endophilins results in the accumulation of clathrin-coated vesicles, but not of clathrin-coated pits, at synapses. The absence of endophilin impairs but does not abolish synaptic transmission and results in perinatal lethality, whereas partial endophilin absence causes severe neurological defects, including epilepsy and neurodegeneration. Our data support a model in which endophilin recruitment to coated pit necks, because of its curvature-sensing properties, primes vesicle buds for subsequent uncoating after membrane fission, without being critically required for the fission reaction itself.
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Program in Cellular Neuroscience, Neurodegeneration, and Repair, Kavli Institute for Neuroscience, Yale University School of Medicine, New Haven, CT 06519, USA.
authors.forename: Ira
authors.initials: I
authors.lastname: Milosevic
authors.forename: Silvia
authors.initials: S
authors.lastname: Giovedi
authors.forename: Xuelin
authors.initials: X
authors.lastname: Lou
authors.forename: Andrea
authors.initials: A
authors.lastname: Raimondi
authors.forename: Chiara
authors.initials: C
authors.lastname: Collesi
authors.forename: Hongying
authors.initials: H
authors.lastname: Shen
authors.forename: Summer
authors.initials: S
authors.lastname: Paradise
authors.forename: Eileen
authors.initials: E
authors.lastname: O'Toole
authors.forename: Shawn
authors.initials: S
authors.lastname: Ferguson
authors.forename: Ottavio
authors.initials: O
authors.lastname: Cremona
authors.forename: Pietro
authors.initials: P
authors.lastname: De Camilli
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000172:deficiency / Q000378:metabolism*; D000818:Animals; D002455:Cell Division / Q000235:genetics; D002462:Cell Membrane / Q000235:genetics / Q000648:ultrastructure; D002966:Clathrin / Q000378:metabolism* / Q000648:ultrastructure; D022163:Clathrin-Coated Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D003034:Coated Pits, Cell-Membrane / Q000378:metabolism* / Q000648:ultrastructure; D047908:Intracellular Signaling Peptides and Proteins / Q000172:deficiency / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008959:Models, Neurological; D021381:Protein Transport / Q000235:genetics; D051381:Rats; D013569:Synapses / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002966:Clathrin; D047908:Intracellular Signaling Peptides and Proteins; C498434:Sh3gl2 protein, mouse; C528010:endophilin II, mouse
references.citation: J Biol Chem. 1997 Mar 28;272(13):8710-6
references.pmid: 9079704
references.citation: Cell. 2009 Apr 17;137(2):191-6
references.pmid: 19379681
references.citation: Dev Cell. 2009 Dec;17(6):811-22
references.pmid: 20059951
references.citation: Proc Natl Acad Sci U S A. 2006 Dec 19;103(51):19332-7
references.pmid: 17158794
references.citation: J Cell Sci. 2011 Jan 1;124(Pt 1):133-43
references.pmid: 21172823
references.citation: Nature. 2006 Oct 12;443(7112):651-7
references.pmid: 17035995
references.citation: Cell. 2002 Apr 5;109(1):101-12
references.pmid: 11955450
references.citation: Neuron. 2003 Nov 13;40(4):749-62
references.pmid: 14622579
references.citation: Structure. 2010 Sep 8;18(9):1191-8
references.pmid: 20826345
references.citation: Curr Opin Cell Biol. 2006 Aug;18(4):386-94
references.pmid: 16782321
references.citation: Neuron. 2000 Aug;27(2):301-12
references.pmid: 10985350
references.citation: Neuron. 1999 Sep;24(1):143-54
references.pmid: 10677033
references.citation: Hum Mol Genet. 2005 Oct 1;14(19):2893-909
references.pmid: 16115810
references.citation: Cell. 2005 Nov 4;123(3):521-33
references.pmid: 16269341
references.citation: Cell. 1999 Oct 15;99(2):179-88
references.pmid: 10535736
references.citation: Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8569-74
references.pmid: 9238017
references.citation: PLoS Biol. 2009 Sep;7(9):e1000204
references.pmid: 19787029
references.citation: Cell. 2010 Oct 29;143(3):430-41
references.pmid: 21029864
references.citation: FEBS Lett. 2010 May 3;584(9):1848-55
references.pmid: 20122931
references.citation: EMBO J. 2006 Jun 21;25(12):2898-910
references.pmid: 16763559
references.citation: Proc Natl Acad Sci U S A. 2004 Mar 16;101(11):3833-8
references.pmid: 15007177
references.citation: Science. 2004 Jan 23;303(5657):495-9
references.pmid: 14645856
references.citation: J Cell Biol. 1973 May;57(2):315-44
references.pmid: 4348786
references.citation: Curr Protoc Cell Biol. 2005 Apr;Chapter 3:Unit 3.13
references.pmid: 18228473
references.citation: Proc Natl Acad Sci U S A. 2006 Jul 5;103(27):10265-10270
references.pmid: 16798879
references.citation: Neuron. 2006 Jul 6;51(1):71-84
references.pmid: 16815333
references.citation: Science. 2007 Apr 27;316(5824):570-4
references.pmid: 17463283
references.citation: Biophys J. 2009 Nov 18;97(10):2746-53
references.pmid: 19917228
references.citation: Annu Rev Cell Dev Biol. 2009;25:133-60
references.pmid: 19575674
references.citation: J Biol Chem. 2001 Nov 2;276(44):40424-30
references.pmid: 11518713
references.citation: Nat Cell Biol. 2000 Apr;2(4):197-204
references.pmid: 10783237
references.citation: Cell. 2005 Oct 21;123(2):305-20
references.pmid: 16239147
references.citation: Mol Cell. 2009 Dec 25;36(6):1034-47
references.pmid: 20064468
references.citation: Proc Natl Acad Sci U S A. 2010 Mar 2;107(9):4412-7
references.pmid: 20160091
references.citation: Proc Natl Acad Sci U S A. 2008 Feb 12;105(6):2175-80
references.pmid: 18250322
references.citation: Neuron. 2003 Nov 13;40(4):733-48
references.pmid: 14622578
references.citation: Nat Rev Mol Cell Biol. 2009 Sep;10(9):583-96
references.pmid: 19696796
references.citation: J Cell Biol. 2001 Oct 15;155(2):193-200
references.pmid: 11604418
references.citation: Dev Cell. 2011 Feb 15;20(2):206-18
references.pmid: 21316588
references.citation: Neuron. 2011 Jun 23;70(6):1100-14
references.pmid: 21689597
references.citation: Neuron. 2011 Mar 24;69(6):1147-59
references.pmid: 21435559
references.citation: Neuron. 2007 Dec 20;56(6):1004-18
references.pmid: 18093523