Re-examining how Munc13-1 facilitates opening of syntaxin-1

Magdalena Magdziarek, Agnieszka A. Bolembach, Karolina P. Stepien, Bradley Quade, Xiaoxia Liu, Josep Rizo

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Munc13-1 is crucial for neurotransmitter release and, together with Munc18-1, orchestrates assembly of the neuronal SNARE complex formed by syntaxin-1, SNAP-25, and synaptobrevin. Assembly starts with syntaxin-1 folded into a self-inhibited closed conformation that binds to Munc18-1. Munc13-1 is believed to catalyze the opening of syntaxin-1 to facilitate SNARE complex formation. However, different types of Munc13-1-syntaxin-1 interactions have been reported to underlie this activity, and the critical nature of Munc13-1 for release may arise because of its key role in bridging the vesicle and plasma membranes. To shed light into the mechanism of action of Munc13-1, we have used NMR spectroscopy, SNARE complex assembly experiments, and liposome fusion assays. We show that point mutations in a linker region of syntaxin-1 that forms intrinsic part of the closed conformation strongly impair stimulation of SNARE complex assembly and liposome fusion mediated by Munc13-1 fragments, even though binding of this linker region to Munc13-1 is barely detectable. Conversely, the syntaxin-1 SNARE motif clearly binds to Munc13-1, but a mutation that disrupts this interaction does not affect SNARE complex assembly or liposome fusion. We also show that Munc13-1 cannot be replaced by an artificial tethering factor to mediate liposome fusion. Overall, these results emphasize how very weak interactions can play fundamental roles in promoting conformational transitions and strongly support a model whereby the critical nature of Munc13-1 for neurotransmitter release arises not only from its ability to bridge two membranes but also from an active role in opening syntaxin-1 via interactions with the linker.

Original languageEnglish (US)
Pages (from-to)1440-1458
Number of pages19
JournalProtein Science
Volume29
Issue number6
DOIs
StatePublished - Jun 1 2020

Keywords

  • Munc13
  • Munc18
  • SNAREs
  • conformational transition
  • membrane fusion
  • neurotransmitter release
  • syntaxin-1

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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