Tyrosine phosphorylation of Munc18-1 inhibits synaptic transmission by preventing SNARE assembly

Marieke Meijer, Bernhard Dörr, Hanna C.A. Lammertse, Chrysanthi Blithikioti, Jan R.T. van Weering, Ruud F.G. Toonen, Thomas H. Söllner, Matthijs Verhage*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review


Tyrosine kinases are important regulators of synaptic strength. Here, we describe a key component of the synaptic vesicle release machinery, Munc18-1, as a phosphorylation target for neuronal Src family kinases (SFKs). Phosphomimetic Y473D mutation of a SFK phosphorylation site previously identified by brain phospho-proteomics abolished the stimulatory effect of Munc18-1 on SNARE complex formation (“SNARE-templating”) and membrane fusion in vitro. Furthermore, priming but not docking of synaptic vesicles was disrupted in hippocampal munc18-1-null neurons expressing Munc18-1Y473D. Synaptic transmission was temporarily restored by high-frequency stimulation, as well as by a Munc18-1 mutation that results in helix 12 extension, a critical conformational step in vesicle priming. On the other hand, expression of non-phosphorylatable Munc18-1 supported normal synaptic transmission. We propose that SFK-dependent Munc18-1 phosphorylation may constitute a potent, previously unknown mechanism to shut down synaptic transmission, via direct occlusion of a Synaptobrevin/VAMP2 binding groove and subsequent hindrance of conformational changes in domain 3a responsible for vesicle priming. This would strongly interfere with the essential post-docking SNARE-templating role of Munc18-1, resulting in a largely abolished pool of releasable synaptic vesicles.

Original languageEnglish
Pages (from-to)300-320
Number of pages21
JournalEMBO Journal
Issue number2
Early online date17 Nov 2017
Publication statusPublished - 17 Jan 2018


We thank Robbert Zalm for virus production, Desiree Schut for primary culture preparation, Ingrid Saarloos for cloning and performing in vitro kinase assays, and Jurjen Broeke and Emmeke Aarts for expert help with analysis. We would like to thank Jörg Malsam and Kerstin Rink for contributing purified proteins to this study. We thank Christiaan van der Meer for animal breeding and maintenance. Rien Dekker is acknowledged for performing electron microscopy. This work was supported by the EU (EUSynapse project 019055 to MV, EUROSPIN project HEALTH-F2-2009-241498 to MV, HEALTH-F2-2009-242167 SynSys project to MV, and ERC Advanced Grant 322966 to MV), the Netherlands Organization for Scientific Research, NWO (Pionier/VICI 900-01-001 and ZonMW 903-42-095 to MV), the NeuroBsik Mouse Phenomics Consortium (BSIK03053), and the German Research Foundation (DFG) (SFB/TRR 83 to BD and THS).

FundersFunder number
Netherlands Organization for Scientific Research
NeuroBsik Mouse Phenomics ConsortiumBSIK03053
Seventh Framework Programme241498, 242167, 322966
European Commission019055, HEALTH-F2-2009-241498
European Research Council
Deutsche ForschungsgemeinschaftSFB/TRR 83
Nederlandse Organisatie voor Wetenschappelijk OnderzoekZonMW 903-42-095, Pionier/VICI 900-01-001


    • Munc18-1
    • priming
    • SNARE
    • Src
    • synaptic transmission


    Dive into the research topics of 'Tyrosine phosphorylation of Munc18-1 inhibits synaptic transmission by preventing SNARE assembly'. Together they form a unique fingerprint.

    Cite this