title: Ca2+-Calmodulin regulates SNARE assembly and spontaneous neurotransmitter release via v-ATPase subunit V0a1.
abstract: Most chemical neurotransmission occurs through Ca(2+)-dependent evoked or spontaneous vesicle exocytosis. In both cases, Ca(2+) sensing is thought to occur shortly before exocytosis. In this paper, we provide evidence that the Ca(2+) dependence of spontaneous vesicle release may partly result from an earlier requirement of Ca(2+) for the assembly of soluble N-ethylmaleimide-sensitive fusion attachment protein receptor (SNARE) complexes. We show that the neuronal vacuolar-type H(+)-adenosine triphosphatase V0 subunit a1 (V100) can regulate the formation of SNARE complexes in a Ca(2+)-Calmodulin (CaM)-dependent manner. Ca(2+)-CaM regulation of V100 is not required for vesicle acidification. Specific disruption of the Ca(2+)-dependent regulation of V100 by CaM led to a >90% loss of spontaneous release but only had a mild effect on evoked release at Drosophila melanogaster embryo neuromuscular junctions. Our data suggest that Ca(2+)-CaM regulation of V100 may control SNARE complex assembly for a subset of synaptic vesicles that sustain spontaneous release.
authors.affiliation: Department of Physiology and 2 Green Center Division for Systems Biology, University of Texas Southwestern Medical Center, Dallas, TX 75390.
authors.forename: Dong
authors.initials: D
authors.lastname: Wang
authors.forename: Daniel
authors.initials: D
authors.lastname: Epstein
authors.forename: Ossama
authors.initials: O
authors.lastname: Khalaf
authors.forename: Sankaranarayanan
authors.initials: S
authors.lastname: Srinivasan
authors.forename: W Ryan
authors.initials: WR
authors.lastname: Williamson
authors.forename: Amir
authors.initials: A
authors.lastname: Fayyazuddin
authors.forename: Florante A
authors.initials: FA
authors.lastname: Quiocho
authors.forename: P Robin
authors.initials: PR
authors.lastname: Hiesinger
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000378:metabolism*; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000201:enzymology* / Q000235:genetics; D004558:Electric Stimulation; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000201:enzymology; D046912:Multiprotein Complexes; D009469:Neuromuscular Junction / Q000201:enzymology*; D011485:Protein Binding; D021122:Protein Subunits; D050765:Qa-SNARE Proteins / Q000235:genetics / Q000378:metabolism*; D009435:Synaptic Transmission; D013572:Synaptic Vesicles / Q000201:enzymology*; D013997:Time Factors; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism*
chemical_list: D002147:Calmodulin; D029721:Drosophila Proteins; D046912:Multiprotein Complexes; D021122:Protein Subunits; D050765:Qa-SNARE Proteins; C578392:syntaxin, Drosophila; D025262:Vacuolar Proton-Translocating ATPases; C550849:Vha100-1 protein, Drosophila; D002118:Calcium
references.citation: Nature. 1982 Jan 14;295(5845):154-5
references.pmid: 7035964
references.citation: J Comp Physiol A. 1994 Aug;175(2):179-91
references.pmid: 8071894
references.citation: Neuron. 2010 Jul 29;67(2):268-79
references.pmid: 20670834
references.citation: Cell Calcium. 1981 Aug;2(4):365-85
references.pmid: 6122506
references.citation: Genetics. 2007 Feb;175(2):709-24
references.pmid: 17179093
references.citation: Neuron. 2013 Jul 10;79(1):82-96
references.pmid: 23770256
references.citation: Cell. 2005 May 20;121(4):607-620
references.pmid: 15907473
references.citation: Nat Neurosci. 2009 Jun;12(6):759-66
references.pmid: 19412166
references.citation: Nature. 2001 Feb 1;409(6820):581-8
references.pmid: 11214310
references.citation: Science. 2008 Sep 12;321(5895):1507-10
references.pmid: 18703708
references.citation: Comput Appl Biosci. 1993 Apr;9(2):197-9
references.pmid: 8481823
references.citation: Cell. 2008 May 30;133(5):916-27
references.pmid: 18510934
references.citation: J Neurosci. 2013 Oct 30;33(44):17253-63
references.pmid: 24174659
references.citation: J Neurosci. 2010 Mar 17;30(11):4132-42
references.pmid: 20237283
references.citation: J Biol Chem. 2010 Jul 30;285(31):23665-75
references.pmid: 20519509
references.citation: Nature. 2007 Nov 29;450(7170):676-82
references.pmid: 18046404
references.citation: Nat Struct Mol Biol. 2008 Jul;15(7):665-74
references.pmid: 18618940
references.citation: Neuron. 1995 Sep;15(3):663-73
references.pmid: 7546745
references.citation: J Cell Biol. 2013 Oct 28;203(2):171-3
references.pmid: 24165933
references.citation: Neuron. 2002 Oct 24;36(3):463-74
references.pmid: 12408848
references.citation: J Biol Chem. 2004 Jan 2;279(1):644-51
references.pmid: 14551199
references.citation: J Cell Biol. 2010 May 31;189(5):885-99
references.pmid: 20513768
references.citation: J Biol Chem. 1996 Jan 26;271(4):2193-8
references.pmid: 8567678
references.citation: J Neurosci. 2003 Nov 19;23(33):10732-44
references.pmid: 14627659
references.citation: EMBO J. 2011 Sep 20;30(20):4126-41
references.pmid: 21934648
references.citation: J Neurosci. 2001 Jun 15;21(12):4215-24
references.pmid: 11404407
references.citation: EMBO J. 2013 Jan 9;32(1):159-71
references.pmid: 23188083
references.citation: J Vis Exp. 2009 May 21;(27):
references.pmid: 19461578
references.citation: J Vis Exp. 2009 May 20;(27):
references.pmid: 19488027
references.citation: Curr Opin Neurobiol. 2005 Jun;15(3):266-74
references.pmid: 15919191
references.citation: Dev Cell. 2013 Nov 25;27(4):462-8
references.pmid: 24286827
references.citation: J Cell Biol. 2010 Oct 4;191(1):187-97
references.pmid: 20921140
references.citation: Genetics. 2012 Jun;191(2):461-75
references.pmid: 22426883
references.citation: Nat Rev Mol Cell Biol. 2002 Feb;3(2):94-103
references.pmid: 11836511
references.citation: J Biol Chem. 1991 Feb 25;266(6):3877-81
references.pmid: 1704894
references.citation: Cell. 2011 Oct 28;147(3):666-77
references.pmid: 22036572
references.citation: Science. 2009 Jan 23;323(5913):474-7
references.pmid: 19164740
references.citation: Mol Biol Cell. 2011 Sep;22(18):3394-409
references.pmid: 21795392
references.citation: Neuron. 2011 Apr 28;70(2):244-51
references.pmid: 21521611
references.citation: Anal Biochem. 2008 Jun 15;377(2):209-17
references.pmid: 18405656
references.citation: Neuron. 2007 May 24;54(4):567-81
references.pmid: 17521570
references.citation: Proc Natl Acad Sci U S A. 2010 Aug 17;107(33):14869-74
references.pmid: 20679236
references.citation: Nature. 2012 Oct 11;490(7419):201-7
references.pmid: 23060190
references.citation: Neuron. 2005 Feb 17;45(4):563-73
references.pmid: 15721242
references.citation: Curr Opin Neurobiol. 2011 Apr;21(2):275-82
references.pmid: 21334193
references.citation: Nature. 2001 Jul 19;412(6844):338-41
references.pmid: 11460165
references.citation: J Vis Exp. 2010 Mar 15;(37):
references.pmid: 20231817
references.citation: J Cell Sci. 2003 Dec 1;116(Pt 23):4751-62
references.pmid: 14600261
references.citation: J Cell Biol. 2013 Oct 28;203(2):283-98
references.pmid: 24165939
references.citation: Science. 2010 Mar 26;327(5973):1614-8
references.pmid: 20150444
references.citation: J Biol Chem. 2008 Jan 4;283(1):294-300
references.pmid: 17933871
references.citation: Neuron. 1999 Jul;23(3):593-605
references.pmid: 10433270