title: A role for an Hsp70 nucleotide exchange factor in the regulation of synaptic vesicle endocytosis.
abstract: Neurotransmission requires a continuously available pool of synaptic vesicles (SVs) that can fuse with the plasma membrane and release their neurotransmitter contents upon stimulation. After fusion, SV membranes and membrane proteins are retrieved from the presynaptic plasma membrane by clathrin-mediated endocytosis. After the internalization of a clathrin-coated vesicle, the vesicle must uncoat to replenish the pool of SVs. Clathrin-coated vesicle uncoating requires ATP and is mediated by the ubiquitous molecular chaperone Hsc70. In vitro, depolymerized clathrin forms a stable complex with Hsc70*ADP. This complex can be dissociated by nucleotide exchange factors (NEFs) that release ADP from Hsc70, allowing ATP to bind and induce disruption of the clathrin:Hsc70 association. Whether NEFs generally play similar roles in vesicle trafficking in vivo and whether they play such roles in SV endocytosis in particular is unknown. To address this question, we used information from recent structural and mechanistic studies of Hsp70:NEF and Hsp70:co-chaperone interactions to design a NEF inhibitor. Using acute perturbations at giant reticulospinal synapses of the sea lamprey (Petromyzon marinus), we found that this NEF inhibitor inhibited SV endocytosis. When this inhibitor was mutated so that it could no longer bind and inhibit Hsp110 (a NEF that we find to be highly abundant in brain cytosol), its ability to inhibit SV endocytosis was eliminated. These observations indicate that the action of a NEF, most likely Hsp110, is normally required during SV trafficking to release clathrin from Hsc70 and make it available for additional rounds of endocytosis.
authors: Eugene Bell Center for Regenerative Biology and Tissue Engineering, Marine Biological Laboratory, Woods Hole, Massachusetts 02543, USA.
authors: Jennifer R
authors: JR
authors: Morgan
authors: Jianwen
authors: J
authors: Jiang
authors: Paul A
authors: PA
authors: Oliphint
authors: Suping
authors: S
authors: Jin
authors: Luis E
authors: LE
authors: Gimenez
authors: David J
authors: DJ
authors: Busch
authors: Andrea E
authors: AE
authors: Foldes
authors: Yue
authors: Y
authors: Zhuo
authors: Rui
authors: R
authors: Sousa
authors: Eileen M
authors: EM
authors: Lafer
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D034021:Auxilins / Q000494:pharmacology; D001921:Brain / Q000166:cytology; D002417:Cattle; D002966:Clathrin / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D005260:Female; D020662:Guanine Nucleotide Exchange Factors / Q000378:metabolism; D050957:HSP110 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007798:Lampreys; D007814:Larva; D008297:Male; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D013116:Spinal Cord / Q000166:cytology; D013572:Synaptic Vesicles / Q000502:physiology*
chemical_list: D034021:Auxilins; D002966:Clathrin; D004268:DNA-Binding Proteins; D020662:Guanine Nucleotide Exchange Factors; D050957:HSP110 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases
references: J Biol Chem. 1994 Apr 29;269(17):12899-907
references: 8175707
references: Cold Spring Harb Perspect Biol. 2012 Sep 01;4(9):a005645
references: 22763746
references: J Mol Biol. 1996 Nov 15;263(5):657-70
references: 8947566
references: EMBO J. 1993 Nov;12(11):4137-44
references: 7900997
references: EMBO J. 2010 Feb 3;29(3):655-65
references: 20033059
references: Traffic. 2007 Sep;8(9):1129-36
references: 17547698
references: Cell. 2008 Jun 13;133(6):1068-79
references: 18555782
references: Nat Struct Biol. 2000 Jul;7(7):586-93
references: 10876246
references: Neuron. 1998 Dec;21(6):1465-75
references: 9883738
references: Mol Cell. 2005 Nov 23;20(4):513-24
references: 16307916
references: J Biol Chem. 2000 Mar 24;275(12):8439-47
references: 10722678
references: Biochemistry. 2006 Dec 19;45(50):15075-84
references: 17154545
references: Nature. 1995 Dec 7;378(6557):632-5
references: 8524399
references: EMBO J. 2006 Jun 7;25(11):2519-28
references: 16688212
references: Nat Struct Mol Biol. 2008 Dec;15(12):1309-17
references: 19029896
references: Neuron. 1999 Sep;24(1):143-54
references: 10677033
references: Cell. 1979 Feb;16(2):303-12
references: 455437
references: J Biol Chem. 2001 Aug 31;276(35):32538-44
references: 11441021
references: J Mol Biol. 2010 Sep 3;401(5):696-707
references: 20624400
references: J Cell Biol. 2004 Oct 11;167(1):43-50
references: 15479735
references: J Biol Chem. 2004 Jul 2;279(27):27957-64
references: 15102842
references: Cell Tissue Res. 2006 Nov;326(2):301-10
references: 16786368
references: Cell. 1986 Jul 4;46(1):5-9
references: 2872968
references: Mol Cell. 2008 Jul 25;31(2):232-43
references: 18550409
references: J Cell Biol. 1987 Nov;105(5):1989-98
references: 2890644
references: Biochem Soc Trans. 2009 Oct;37(Pt 5):1022-6
references: 19754444
references: Neuromolecular Med. 2002;2(2):101-14
references: 12428806
references: Biol Chem. 2005 Aug;386(8):739-44
references: 16201868
references: Science. 2001 Feb 23;291(5508):1553-7
references: 11222862
references: J Biol Chem. 2012 Feb 17;287(8):5661-72
references: 22157767
references: Cell. 1997 Mar 21;88(6):767-76
references: 9118220
references: FEBS Lett. 2006 Jan 9;580(1):168-74
references: 16364315
references: Proc Natl Acad Sci U S A. 2004 Mar 16;101(11):3833-8
references: 15007177
references: J Neurosci. 1999 Dec 1;19(23):10201-12
references: 10575017
references: J Biol Chem. 1993 Jun 15;268(17):12655-62
references: 7685348
references: J Mol Biol. 1975 Sep 5;97(1):93-8
references: 1177317
references: Biochem Soc Trans. 2006 Feb;34(Pt 1):68-72
references: 16417485
references: Neuron. 1998 Sep;21(3):607-16
references: 9768846
references: J Biotechnol. 2002 Jun 13;96(1):13-21
references: 12142139
references: Oncogene. 2003 Aug 7;22(32):4973-82
references: 12902980
references: J Neurocytol. 2003 Jun-Sep;32(5-8):567-89
references: 15034254
references: Mol Biol Cell. 2008 Jan;19(1):405-13
references: 17978091
references: Biochim Biophys Acta. 1999 Jan 18;1444(1):138-42
references: 9931472
references: Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10345-9
references: 7937953
references: J Biol Chem. 1997 Mar 7;272(10):6393-8
references: 9045662
references: J Cell Sci. 2011 Jan 1;124(Pt 1):133-43
references: 21172823
references: Nature. 1995 Mar 9;374(6518):186-90
references: 7877693
references: J Cell Biol. 2001 Feb 5;152(3):607-20
references: 11157986
references: Cell. 2004 Nov 24;119(5):653-65
references: 15550247
references: Cell. 2011 Aug 5;146(3):471-84
references: 21816279
references: Proc Natl Acad Sci U S A. 1982 Oct;79(19):5881-5
references: 6964393
references: J Neurosci. 2000 Dec 1;20(23):8667-76
references: 11102472
references: J Comp Neurol. 2010 Jul 15;518(14):2854-72
references: 20506479
references: Annu Rev Cell Dev Biol. 2009;25:133-60
references: 19575674
references: PLoS One. 2011;6(10):e26319
references: 22022600
references: J Biol Chem. 2003 Aug 29;278(35):33583-92
references: 12807910
references: J Neurosci Res. 1995 May 1;41(1):15-26
references: 7674375
references: Proc Natl Acad Sci U S A. 2010 Mar 2;107(9):4412-7
references: 20160091
references: Mol Cell. 2005 Feb 4;17(3):367-79
references: 15694338
references: Science. 2001 Feb 9;291(5506):1051-5
references: 11161218
references: Structure. 2010 Mar 10;18(3):309-19
references: 20223214
references: J Biol Chem. 1999 Aug 6;274(32):22785-94
references: 10428863
references: Nature. 1995 Jun 8;375(6531):493-7
references: 7777058
references: Biochemistry. 1996 Apr 9;35(14):4636-44
references: 8605215
references: Science. 1997 Apr 11;276(5310):259-63
references: 9092476
references: J Comp Neurol. 2006 Aug 1;497(4):600-9
references: 16739194
references: J Neurosci. 1991 May;11(5):1163-72
references: 1709204
references: Mol Cell. 2007 Nov 9;28(3):422-33
references: 17996706
references: Science. 1997 Apr 18;276(5311):431-5
references: 9103205
references: EMBO J. 1986 Dec 1;5(12):3143-9
references: 3816757
references: Neuron. 2001 Oct 25;32(2):289-300
references: 11683998