title: Comparative studies of Munc18c and Munc18-1 reveal conserved and divergent mechanisms of Sec1/Munc18 proteins.
abstract: Sec1/Munc18 (SM) family proteins are essential for every vesicle fusion pathway. The best-characterized SM protein is the synaptic factor Munc18-1, but it remains unclear whether its functions represent conserved mechanisms of SM proteins or specialized activities in neurotransmitter release. To address this question, we dissected Munc18c, a functionally distinct SM protein involved in nonsynaptic exocytic pathways. We discovered that Munc18c binds to the trans-SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) complex and strongly accelerates the fusion rate. Further analysis suggests that Munc18c recognizes both vesicle-rooted SNARE and target membrane-associated SNAREs, and promotes trans-SNARE zippering at the postdocking stage of the fusion reaction. The stimulation of fusion by Munc18c is specific to its cognate SNARE isoforms. Because Munc18-1 regulates fusion in a similar manner, we conclude that one conserved function of SM proteins is to bind their cognate trans-SNARE complexes and accelerate fusion kinetics. Munc18c also binds syntaxin-4 monomer but does not block target membrane-associated SNARE assembly, in agreement with our observation that six- to eightfold increases in Munc18c expression do not inhibit insulin-stimulated glucose uptake in adipocytes. Thus, the inhibitory "closed" syntaxin binding mode demonstrated for Munc18-1 is not conserved in Munc18c. Unexpectedly, we found that Munc18c recognizes the N-terminal region of the vesicle-rooted SNARE, whereas Munc18-1 requires the C-terminal sequences, suggesting that the architecture of the SNARE/SM complex likely differs across fusion pathways. Together, these comparative studies of two distinct SM proteins reveal conserved as well as divergent mechanisms of SM family proteins in intracellular vesicle fusion.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309, USA.
authors.forename: Haijia
authors.initials: H
authors.lastname: Yu
authors.forename: Shailendra S
authors.initials: SS
authors.lastname: Rathore
authors.forename: Jamie A
authors.initials: JA
authors.lastname: Lopez
authors.forename: Eric M
authors.initials: EM
authors.lastname: Davis
authors.forename: David E
authors.initials: DE
authors.lastname: James
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Martin
authors.forename: Jingshi
authors.initials: J
authors.lastname: Shen
mesh_terms: D005089:Exocytosis; D007700:Kinetics; D008561:Membrane Fusion; D051938:Munc18 Proteins / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D050600:SNARE Proteins / Q000378:metabolism
chemical_list: D051938:Munc18 Proteins; D050600:SNARE Proteins
keywords: exocytosis; membrane fusion; vesicle transport
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