title: Modification of a hydrophobic layer by a point mutation in syntaxin 1A regulates the rate of synaptic vesicle fusion.
abstract: Both constitutive secretion and Ca(2+)-regulated exocytosis require the assembly of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complexes. At present, little is known about how the SNARE complexes mediating these two distinct pathways differ in structure. Using the Drosophila neuromuscular synapse as a model, we show that a mutation modifying a hydrophobic layer in syntaxin 1A regulates the rate of vesicle fusion. Syntaxin 1A molecules share a highly conserved threonine in the C-terminal +7 layer near the transmembrane domain. Mutation of this threonine to isoleucine results in a structural change that more closely resembles those found in syntaxins ascribed to the constitutive secretory pathway. Flies carrying the I254 mutant protein have increased levels of SNARE complexes and dramatically enhanced rate of both constitutive and evoked vesicle fusion. In contrast, overexpression of the T254 wild-type protein in neurons reduces vesicle fusion only in the I254 mutant background. These results are consistent with molecular dynamics simulations of the SNARE core complex, suggesting that T254 serves as an internal brake to dampen SNARE zippering and impede vesicle fusion, whereas I254 favors fusion by enhancing intermolecular interaction within the SNARE core complex.
authors.affiliation: Section of Neurobiology and Institute for Neuroscience, University of Texas at Austin, Austin, Texas, United States of America.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Lagow
authors.forename: Hong
authors.initials: H
authors.lastname: Bao
authors.forename: Evan N
authors.initials: EN
authors.lastname: Cohen
authors.forename: Richard W
authors.initials: RW
authors.lastname: Daniels
authors.forename: Aleksej
authors.initials: A
authors.lastname: Zuzek
authors.forename: Wade H
authors.initials: WH
authors.lastname: Williams
authors.forename: Gregory T
authors.initials: GT
authors.lastname: Macleod
authors.forename: R Bryan
authors.initials: RB
authors.lastname: Sutton
authors.forename: Bing
authors.initials: B
authors.lastname: Zhang
mesh_terms: D000200:Action Potentials; D000818:Animals; D004330:Drosophila; D017354:Point Mutation; D050600:SNARE Proteins / Q000502:physiology; D013572:Synaptic Vesicles / Q000502:physiology*; D050827:Syntaxin 1 / Q000235:genetics*
chemical_list: D050600:SNARE Proteins; D050827:Syntaxin 1