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PMID: 24120845 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Lipid-anchored SNAREs lacking transmembrane regions fully support membrane fusion during neurotransmitter release.

Neuron ·Vol. 80 ·No. 2 ·2013-10-16 ·Pages 470-83

Zhou P, Bacaj T, Yang X, Pang ZP, Südhof TC

Abstract

Synaptic vesicle fusion during neurotransmitter release is mediated by assembly of SNARE- and SM-protein complexes composed of syntaxin-1, SNAP-25, synaptobrevin-2/VAMP2, and Munc18-1. Current models suggest that SNARE-complex assembly catalyzes membrane fusion by pulling the transmembrane regions (TMRs) of SNARE proteins together, thus allowing their TMRs to form a fusion pore. These models are consistent with the requirement for TMRs in viral fusion proteins. However, the role of the SNARE TMRs in synaptic vesicle fusion has not yet been tested physiologically. Here, we examined whether synaptic SNAREs require TMRs for catalysis of synaptic vesicle fusion, which was monitored electrophysiologically at millisecond time resolution. Surprisingly, we find that both lipid-anchored syntaxin-1 and lipid-anchored synaptobrevin-2 lacking TMRs efficiently promoted spontaneous and Ca(2+)-triggered membrane fusion. Our data suggest that SNARE proteins function during fusion primarily as force generators, consistent with the notion that forcing lipid membranes close together suffices to induce membrane fusion.

MeSH Terms
Animals Cells, Cultured Inhibitory Postsynaptic Potentials/physiology Membrane Fusion/physiology Membrane Lipids/metabolism Mice Mice, Knockout Mice, Transgenic Neurons/physiology Neurotransmitter Agents/metabolism SNARE Proteins/chemistry,genetics,metabolism Synaptic Transmission/physiology Synaptic Vesicles/metabolism Syntaxin 1/genetics,physiology Vesicle-Associated Membrane Protein 2/genetics,physiology
Chemicals
Membrane Lipids Neurotransmitter Agents SNARE Proteins Syntaxin 1 Vesicle-Associated Membrane Protein 2
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zhou Peng
Department of Molecular and Cellular Physiology and Howard Hughes Medical Institute, Stanford University School of Medicine, 265 Campus Drive, Stanford, CA 94305, USA.
Bacaj Taulant
Yang Xiaofei
Pang Zhiping P
Südhof Thomas C
References (53)
53 references, click to expand
  1. The dynamin-like GTPase Sey1p mediates homotypic ER fusion in S. cerevisiae.
    J Cell Biol. 2012 Apr 16;197(2):209-17 PMID: 22508509
  2. GPI-anchored influenza hemagglutinin induces hemifusion to both red blood cell and planar bilayer membranes.
    J Cell Biol. 1995 Nov;131(3):679-91 PMID: 7593189
  3. Membrane-proximal tryptophans of synaptobrevin II stabilize priming of secretory vesicles.
    J Neurosci. 2012 Nov 7;32(45):15983-97 PMID: 23136435
  4. Self-interaction of a SNARE transmembrane domain promotes the hemifusion-to-fusion transition.
    J Mol Biol. 2006 Dec 15;364(5):1048-60 PMID: 17054985
  5. Control of fusion pore dynamics during exocytosis by Munc18.
    Science. 2001 Feb 2;291(5505):875-8 PMID: 11157167
  6. Structural determinants of synaptobrevin 2 function in synaptic vesicle fusion.
    J Neurosci. 2006 Jun 21;26(25):6668-76 PMID: 16793874
  7. Dual modes of Munc18-1/SNARE interactions are coupled by functionally critical binding to syntaxin-1 N terminus.
    J Neurosci. 2007 Nov 7;27(45):12147-55 PMID: 17989281
  8. Syntaxin N-terminal peptide motif is an initiation factor for the assembly of the SNARE-Sec1/Munc18 membrane fusion complex.
    Proc Natl Acad Sci U S A. 2010 Dec 28;107(52):22399-406 PMID: 21139055
  9. Synaptic assembly of the brain in the absence of neurotransmitter secretion.
    Science. 2000 Feb 4;287(5454):864-9 PMID: 10657302
  10. A fast mode of membrane fusion dependent on tight SNARE zippering.
    J Neurosci. 2008 Aug 20;28(34):8470-6 PMID: 18716205
  11. A class of dynamin-like GTPases involved in the generation of the tubular ER network.
    Cell. 2009 Aug 7;138(3):549-61 PMID: 19665976
  12. Intracellular and viral membrane fusion: a uniting mechanism.
    Curr Opin Cell Biol. 2004 Aug;16(4):429-35 PMID: 15261676
  13. Caught in the act: visualization of SNARE-mediated fusion events in molecular detail.
    Chembiochem. 2011 May 2;12(7):1049-55 PMID: 21433241
  14. Role of the synaptobrevin C terminus in fusion pore formation.
    Proc Natl Acad Sci U S A. 2010 Oct 26;107(43):18463-8 PMID: 20937897
  15. Membrane fusion.
    Cell. 2003 Feb 21;112(4):519-33 PMID: 12600315
  16. Complexin clamps asynchronous release by blocking a secondary Ca(2+) sensor via its accessory α helix.
    Neuron. 2010 Dec 9;68(5):907-20 PMID: 21145004
  17. The dynamin-related GTPase, Mgm1p, is an intermembrane space protein required for maintenance of fusion competent mitochondria.
    J Cell Biol. 2000 Oct 16;151(2):341-52 PMID: 11038181
  18. Geranylgeranylated SNAREs are dominant inhibitors of membrane fusion.
    J Cell Biol. 2000 Oct 16;151(2):453-66 PMID: 11038190
  19. Synaptic vesicle fusion.
    Nat Struct Mol Biol. 2008 Jul;15(7):665-74 PMID: 18618940
  20. Lipid-anchored influenza hemagglutinin promotes hemifusion, not complete fusion.
    Cell. 1994 Jan 28;76(2):383-91 PMID: 8293471
  21. One SNARE complex is sufficient for membrane fusion.
    Nat Struct Mol Biol. 2010 Mar;17(3):358-64 PMID: 20139985
  22. rab3A attachment to the synaptic vesicle membrane mediated by a conserved polyisoprenylated carboxy-terminal sequence.
    Neuron. 1991 Jul;7(1):101-9 PMID: 1648935
  23. Selective activation of cognate SNAREpins by Sec1/Munc18 proteins.
    Cell. 2007 Jan 12;128(1):183-95 PMID: 17218264
  24. v-SNARE actions during Ca(2+)-triggered exocytosis.
    Cell. 2007 Oct 19;131(2):351-63 PMID: 17956735
  25. SNARE function analyzed in synaptobrevin/VAMP knockout mice.
    Science. 2001 Nov 2;294(5544):1117-22 PMID: 11691998
  26. Supramolecular SNARE assembly precedes hemifusion in SNARE-mediated membrane fusion.
    Nat Struct Mol Biol. 2008 Jul;15(7):700-6 PMID: 18552827
  27. SNARE force synchronizes synaptic vesicle fusion and controls the kinetics of quantal synaptic transmission.
    J Neurosci. 2010 Aug 4;30(31):10272-81 PMID: 20685972
  28. Transmembrane segments of syntaxin line the fusion pore of Ca2+-triggered exocytosis.
    Science. 2004 Apr 9;304(5668):289-92 PMID: 15016962
  29. Helical extension of the neuronal SNARE complex into the membrane.
    Nature. 2009 Jul 23;460(7254):525-8 PMID: 19571812
  30. Hemifusion in SNARE-mediated membrane fusion.
    Nat Struct Mol Biol. 2005 May;12(5):417-22 PMID: 15821745
  31. Fast vesicle fusion in living cells requires at least three SNARE complexes.
    Science. 2010 Oct 22;330(6003):502-5 PMID: 20847232
  32. Two synaptobrevin molecules are sufficient for vesicle fusion in central nervous system synapses.
    Proc Natl Acad Sci U S A. 2011 Aug 23;108(34):14318-23 PMID: 21844343
  33. Munc18-1 binds directly to the neuronal SNARE complex.
    Proc Natl Acad Sci U S A. 2007 Feb 20;104(8):2697-702 PMID: 17301226
  34. A conserved molecular pathway mediates myoblast fusion in insects and vertebrates.
    Nat Genet. 2007 Jun;39(6):781-6 PMID: 17529975
  35. Transmembrane-domain determinants for SNARE-mediated membrane fusion.
    J Cell Sci. 2010 Jul 15;123(Pt 14):2473-80 PMID: 20571052
  36. The synaptic vesicle cycle: a cascade of protein-protein interactions.
    Nature. 1995 Jun 22;375(6533):645-53 PMID: 7791897
  37. Autonomous function of synaptotagmin 1 in triggering synchronous release independent of asynchronous release.
    Neuron. 2005 Nov 23;48(4):547-54 PMID: 16301172
  38. Syntaxin-1 N-peptide and Habc-domain perform distinct essential functions in synaptic vesicle fusion.
    EMBO J. 2013 Jan 9;32(1):159-71 PMID: 23188083
  39. Definition of the readily releasable pool of vesicles at hippocampal synapses.
    Neuron. 1996 Jun;16(6):1197-207 PMID: 8663996
  40. Reconstitution of the vital functions of Munc18 and Munc13 in neurotransmitter release.
    Science. 2013 Jan 25;339(6118):421-5 PMID: 23258414
  41. A conserved membrane-spanning amino acid motif drives homomeric and supports heteromeric assembly of presynaptic SNARE proteins.
    J Biol Chem. 2000 Jun 9;275(23):17481-7 PMID: 10764817
  42. Membrane fusion: grappling with SNARE and SM proteins.
    Science. 2009 Jan 23;323(5913):474-7 PMID: 19164740
  43. An isolated pool of vesicles recycles at rest and drives spontaneous neurotransmission.
    Neuron. 2005 Feb 17;45(4):563-73 PMID: 15721242
  44. LTP requires a unique postsynaptic SNARE fusion machinery.
    Neuron. 2013 Feb 6;77(3):542-58 PMID: 23395379
  45. Calmodulin suppresses synaptotagmin-2 transcription in cortical neurons.
    J Biol Chem. 2010 Oct 29;285(44):33930-9 PMID: 20729199
  46. Sec18p and Vam7p remodel trans-SNARE complexes to permit a lipid-anchored R-SNARE to support yeast vacuole fusion.
    EMBO J. 2007 Dec 12;26(24):4935-45 PMID: 18007597
  47. SNARE proteins: one to fuse and three to keep the nascent fusion pore open.
    Science. 2012 Mar 16;335(6074):1355-9 PMID: 22422984
  48. A stable interaction between syntaxin 1a and synaptobrevin 2 mediated by their transmembrane domains.
    FEBS Lett. 1999 Mar 5;446(1):40-4 PMID: 10100611
  49. Conformational switch of syntaxin-1 controls synaptic vesicle fusion.
    Science. 2008 Sep 12;321(5895):1507-10 PMID: 18703708
  50. Complexin regulates the closure of the fusion pore during regulated vesicle exocytosis.
    J Biol Chem. 2002 May 24;277(21):18249-52 PMID: 11929859
  51. A lipid-anchored SNARE supports membrane fusion.
    Proc Natl Acad Sci U S A. 2011 Oct 18;108(42):17325-30 PMID: 21987819
  52. Conflicting views on the membrane fusion machinery and the fusion pore.
    Annu Rev Cell Dev Biol. 2009;25:513-37 PMID: 19575641
  53. Complexin controls the force transfer from SNARE complexes to membranes in fusion.
    Science. 2009 Jan 23;323(5913):516-21 PMID: 19164751
Article Info
Journal
Neuron
Abbr.
Neuron
ISSN
1097-4199
DOI
10.1016/j.neuron.2013.09.010
Published
2013-10-16
Epub
2013-00-10
Pages
470-83
Language
English
Region
United States
NLM ID
8809320
PMCID
PMC3872166
Subset
IM
Grants
NIMH NIH HHS · R01 MH089054 · United States
NINDS NIH HHS · F32 NS067896 · United States
NINDS NIH HHS · 1F32NS067896 · United States
Howard Hughes Medical Institute · United States
NIMH NIH HHS · P50 MH086403 · United States
NINDS NIH HHS · R01 NS077906 · United States
NIMH NIH HHS · 1R01 MH089054 · United States
Corrections
CommentIn
PMID: 24176637
External Links
PubMed source DOI: 10.1016/j.neuron.2013.09.010 PMC Full Text View journal details
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