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Published online
doi:10.1083/jcb.200812026
The Journal of Cell Biology, Vol. 184, No. 5, 751-764
The Rockefeller University Press, 0021-9525 $30.00
© Deák et al.
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Article

Munc18-1 binding to the neuronal SNARE complex controls synaptic vesicle priming



Ferenc Deák1,2, Yi Xu3,4, Wen-Pin Chang2, Irina Dulubova3,4, Mikhail Khvotchev1,2, Xinran Liu2,5, Thomas C. Südhof1,2,5, and Josep Rizo3,4

1 Howard Hughes Medical Institute, 2 Department of Neuroscience, 3 Department of Biochemistry, 4 Department of Pharmacology, and 5 Department of Molecular Genetics, University of Texas Southwestern Medical Center, Dallas, TX 75390

Correspondence to Thomas C. Südhof: tcs1{at}stanford.edu; or Josep Rizo: jose{at}arnie.swmed.edu

Munc18-1 and soluble NSF attachment protein receptors (SNAREs) are critical for synaptic vesicle fusion. Munc18-1 binds to the SNARE syntaxin-1 folded into a closed conformation and to SNARE complexes containing open syntaxin-1. Understanding which steps in fusion depend on the latter interaction and whether Munc18-1 competes with other factors such as complexins for SNARE complex binding is critical to elucidate the mechanisms involved. In this study, we show that lentiviral expression of Munc18-1 rescues abrogation of release in Munc18-1 knockout mice. We describe point mutations in Munc18-1 that preserve tight binding to closed syntaxin-1 but markedly disrupt Munc18-1 binding to SNARE complexes containing open syntaxin-1. Lentiviral rescue experiments reveal that such disruption selectively impairs synaptic vesicle priming but not Ca2+-triggered fusion of primed vesicles. We also find that Munc18-1 and complexin-1 bind simultaneously to SNARE complexes. These results suggest that Munc18-1 binding to SNARE complexes mediates synaptic vesicle priming and that the resulting primed state involves a Munc18-1–SNARE–complexin macromolecular assembly that is poised for Ca2+ triggering of fusion.

F. Deák and Y. Xu contributed equally to this paper.

F. Deák's present address is Dept. of Neuroscience, Mayo Clinic, Jacksonville, FL 32224.

I. Dulubova's present address is Reata Pharmaceuticals, Inc., Irving, TX 75063.

T.C. Südhof's present address is Dept. of Molecular and Cellular Physiology and Stanford Institute for Neuro-Innovation and Translational Neuroscience, Stanford University School of Medicine, Stanford, CA 94305.

Abbreviations used in this paper: DIV, day in vitro; ITC, isothermal titration calorimetry; KO, knockout; NMR, nuclear magnetic resonance; RRP, readily releasable pool; SM, Sec1/Munc18; SMR, strongest methyl resonance; WT, wild type.

© 2009 Deák et al.
This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).


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