Control of neurotransmitter release by two distinct membrane-binding faces of the Munc13-1 C 1 C 2 B region.

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  • Additional Information
    • Source:
      Publisher: eLife Sciences Publications, Ltd Country of Publication: England NLM ID: 101579614 Publication Model: Electronic Cited Medium: Internet ISSN: 2050-084X (Electronic) Linking ISSN: 2050084X NLM ISO Abbreviation: Elife Subsets: MEDLINE
    • Publication Information:
      Original Publication: Cambridge, UK : eLife Sciences Publications, Ltd., 2012-
    • Subject Terms:
      Intracellular Signaling Peptides and Proteins/*metabolism ; Nerve Tissue Proteins/*metabolism ; Neurotransmitter Agents/*metabolism ; Synaptic Vesicles/*metabolism; Animals ; Biological Transport ; Biophysical Phenomena ; Cell Communication ; Cell Membrane/metabolism ; Cells, Cultured ; Hippocampus/cytology ; Intracellular Signaling Peptides and Proteins/genetics ; Mice, Knockout ; Molecular Dynamics Simulation ; Mutation ; Nerve Tissue Proteins/genetics ; Neurons/metabolism ; Synaptic Transmission ; Mice
    • Abstract:
      Munc13-1 plays a central role in neurotransmitter release through its conserved C-terminal region, which includes a diacyglycerol (DAG)-binding C 1 domain, a Ca 2+ /PIP 2 -binding C 2 B domain, a MUN domain and a C 2 C domain. Munc13-1 was proposed to bridge synaptic vesicles to the plasma membrane through distinct interactions of the C 1 C 2 B region with the plasma membrane: (i) one involving a polybasic face that is expected to yield a perpendicular orientation of Munc13-1 and hinder release; and (ii) another involving the DAG-Ca 2+ -PIP 2 -binding face that is predicted to result in a slanted orientation and facilitate release. Here, we have tested this model and investigated the role of the C 1 C 2 B region in neurotransmitter release. We find that K603E or R769E point mutations in the polybasic face severely impair Ca 2+ -independent liposome bridging and fusion in in vitro reconstitution assays, and synaptic vesicle priming in primary murine hippocampal cultures. A K720E mutation in the polybasic face and a K706E mutation in the C 2 B domain Ca 2+ -binding loops have milder effects in reconstitution assays and do not affect vesicle priming, but enhance or impair Ca 2+ -evoked release, respectively. The phenotypes caused by combining these mutations are dominated by the K603E and R769E mutations. Our results show that the C 1 -C 2 B region of Munc13-1 plays a central role in vesicle priming and support the notion that two distinct faces of this region control neurotransmitter release and short-term presynaptic plasticity.
      Competing Interests: MC, BQ, TT, JX, LS, JR, CR No competing interests declared
      (© 2021, Camacho et al.)
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    • Grant Information:
      R35 NS097333 United States NS NINDS NIH HHS; T32 GM008297 United States GM NIGMS NIH HHS
    • Contributed Indexing:
      Keywords: Munc13; mouse; neuroscience; neurotransmitter release; presynaptic plasticity; synaptic vesicle fusion
    • Accession Number:
      0 (Intracellular Signaling Peptides and Proteins)
      0 (Nerve Tissue Proteins)
      0 (Neurotransmitter Agents)
      0 (Unc13a protein, mouse)
      0 (Unc13b protein, mouse)
    • Publication Date:
      Date Created: 20211115 Date Completed: 20211214 Latest Revision: 20240226
    • Publication Date:
      20240226
    • Accession Number:
      PMC8648301
    • Accession Number:
      10.7554/eLife.72030
    • Accession Number:
      34779770