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© The Rockefeller University Press, 0021-9525/2000//889 $5.00
The Journal of Cell Biology, Volume 149, Number 4, , 2000 889-900

Role of Tetanus Neurotoxin Insensitive Vesicle-Associated Membrane Protein (Ti-Vamp) in Vesicular Transport Mediating Neurite Outgrowth

^a Group of Membrane Traffic and Neuronal Plasticity, INSERM U536
^b Group of Morphogenesis and Cell Signaling, CNRS UMR144, Institut Curie, F-75005 Paris, France
INSERM U536 and CNRS UMR144, Institut Curie, Section de Recherche, 26 rue d'Ulm, 75231 Paris Cédex 05, France.33 142 346 37733 142 346 372

thierry.galli{at}curie.fr

How vesicular transport participates in neurite outgrowth is still poorly understood. Neurite outgrowth is not sensitive to tetanus neurotoxin thus does not involve synaptobrevin-mediated vesicular transport to the plasma membrane of neurons. Tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI-VAMP) is a vesicle-SNARE (soluble N-ethylmaleimide-sensitive fusion protein [NSF] attachment protein [SNAP] receptor), involved in transport to the apical plasma membrane in epithelial cells, a tetanus neurotoxin-resistant pathway. Here we show that TI-VAMP is essential for vesicular transport-mediating neurite outgrowth in staurosporine-differentiated PC12 cells. The NH₂-terminal domain, which precedes the SNARE motif of TI-VAMP, inhibits the association of TI-VAMP with synaptosome-associated protein of 25 kD (SNAP25). Expression of this domain inhibits neurite outgrowth as potently as Botulinum neurotoxin E, which cleaves SNAP25. In contrast, expression of the NH₂-terminal deletion mutant of TI-VAMP increases SNARE complex formation and strongly stimulates neurite outgrowth. These results provide the first functional evidence for the role of TI-VAMP in neurite outgrowth and point to its NH₂-terminal domain as a key regulator in this process.

Key Words: membrane traffic • neurite outgrowth • SNAREs • TI-VAMP • SNAP25

© 2000 The Rockefeller University Press

Abbreviations used in this paper: Nter-TIVAMP, NH₂-terminal domain-TIVAMP; BoNTs, Botulinum NTs; Cyt-TIVAMP, cytosolic domain-TIVAMP; GFP, green fluorescent protein; GFP-Cb, GFP-cellubrevin; GST, glutathione-S-transferase; LAMP1, lysosome-associated membrane protein 1; NSF, N-ethylmaleimide–sensitive fusion protein; NTs, neurotoxins; Nter-TIVAMP, NH₂-terminal domain-TIVAMP; SNAP, NSF attachment protein; SNARE, SNAP receptors; SVs, synaptic vesicles; TeNT, tetanus neurotoxin; TeNT-LC, TeNT light chain; TI-VAMP, TeNT-insensitive VAMP; t-SNARE, target SNARE; VAMP, vesicle-associated membrane protein; v-SNARE, vesicle SNARE.

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