A dual mechanism controlling the localization and function of exocytic v-SNAREs

SNARE [soluble NSF (N-ethylmaleimide-sensitive factor) attachment protein receptor] proteins are essential for membrane fusion but their regulation is not yet fully understood. We have previously shown that the amino-terminal Longin domain of the v-SNARE TI-VAMP (tetanus neurotoxin-insensitive vesicle-associated membrane protein)/VAMP7 plays an inhibitory role in neurite outgrowth. The goal of this study was to investigate the regulation of TI-VAMP as a model of v-SNARE regulation. We show here that the Longin domain (LD) plays a dual role. First, it negatively regulates the ability of TI-VAMP and of a Longin/Synaptobrevin chimera to participate in SNARE complexes. Second, it interacts with the adaptor complex AP-3 and this interaction targets TI-VAMP to late endosomes. Accordingly, in mocha cells lacking AP-3δ, TI-VAMP is retained in an early endosomal compartment. Furthermore, TI-VAMPc, an isoform of TI-VAMP lacking part of the LD, does not interact with AP-3, and therefore is not targeted to late endosomes; however, this shorter LD still inhibits SNARE-complex formation. These findings support a mechanism controlling both localization and function of TI-VAMP through the LD and clathrin adaptors. Moreover, they point to the amino-terminal domains of SNARE proteins as multifunctional modules responsible for the fine tuning of SNARE function.

[1]  Haruki Hasegawa,et al.  Mammalian ykt6 is a neuronal SNARE targeted to a specialized compartment by its profilin-like amino terminal domain. , 2003, Molecular biology of the cell.

[2]  U. Koszinowski,et al.  AP‐1A and AP‐3A Lysosomal Sorting Functions , 2002, Traffic.

[3]  J. Rothman,et al.  The machinery and principles of vesicle transport in the cell , 2002, Nature Medicine.

[4]  I. Dulubova,et al.  The N-terminal Domains of Syntaxin 7 and vti1b Form Three-helix Bundles That Differ in Their Ability to Regulate SNARE Complex Assembly* 210 , 2002, The Journal of Biological Chemistry.

[5]  K. Hahn,et al.  The δ subunit of AP-3 is required for efficient transport of VSV-G from the trans-Golgi network to the cell surface , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Bor Luen Tang,et al.  Early/recycling endosomes-to-TGN transport involves two SNARE complexes and a Rab6 isoform , 2002, The Journal of cell biology.

[7]  R. Scheller,et al.  The Di-leucine Motif of Vesicle-associated Membrane Protein 4 Is Required for Its Localization and AP-1 Binding* , 2001, The Journal of Biological Chemistry.

[8]  R. Kelly,et al.  The Neuronal Form of Adaptor Protein-3 Is Required for Synaptic Vesicle Formation from Endosomes , 2001, The Journal of Neuroscience.

[9]  H. Tochio,et al.  An Autoinhibitory Mechanism for Nonsyntaxin SNARE Proteins Revealed by the Structure of Ykt6p , 2001, Science.

[10]  M. D'Esposito,et al.  Longins: a new evolutionary conserved VAMP family sharing a novel SNARE domain. , 2001, Trends in biochemical sciences.

[11]  W. Weis,et al.  A Novel SNARE N-terminal Domain Revealed by the Crystal Structure of Sec22b* , 2001, The Journal of Biological Chemistry.

[12]  A. Prochiantz,et al.  A Common Exocytotic Mechanism Mediates Axonal and Dendritic Outgrowth , 2001, The Journal of Neuroscience.

[13]  J. Wojcik,et al.  The protein–protein interaction map of Helicobacter pylori , 2001, Nature.

[14]  M. Mercola,et al.  TGF-β Superfamily Signaling and Left-Right Asymmetry , 2001, Science's STKE.

[15]  J. Andersson,et al.  Differential sorting of SNAP-25a and SNAP-25b proteins in neuroblastoma cells. , 2000, European journal of cell biology.

[16]  T. Galli,et al.  Soluble NSF Attachment Protein Receptors (SNAREs) in RBL-2H3 Mast Cells: Functional Role of Syntaxin 4 in Exocytosis and Identification of a Vesicle-Associated Membrane Protein 8-Containing Secretory Compartment1 , 2000, The Journal of Immunology.

[17]  T. Galli,et al.  Role of Tetanus Neurotoxin Insensitive Vesicle-Associated Membrane Protein (Ti-Vamp) in Vesicular Transport Mediating Neurite Outgrowth , 2000, The Journal of cell biology.

[18]  T. Galli,et al.  Subcellular Localization of Tetanus Neurotoxin-Insensitive Vesicle-Associated Membrane Protein (VAMP)/VAMP7 in Neuronal Cells: Evidence for a Novel Membrane Compartment , 1999, The Journal of Neuroscience.

[19]  J. Rothman,et al.  Rapid and efficient fusion of phospholipid vesicles by the alpha-helical core of a SNARE complex in the absence of an N-terminal regulatory domain. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[20]  R. Scheller,et al.  Vamp-7 Mediates Vesicular Transport from Endosomes to Lysosomes , 1999, The Journal of cell biology.

[21]  T. Galli,et al.  Raft association of SNAP receptors acting in apical trafficking in Madin-Darby canine kidney cells. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[22]  C. Burd,et al.  Acidic Di-leucine Motif Essential for AP-3–dependent Sorting and Restriction of the Functional Specificity of the Vam3p Vacuolar t-SNARE , 1998, The Journal of cell biology.

[23]  R. Schekman,et al.  Nucleation of COPII vesicular coat complex by endoplasmic reticulum to Golgi vesicle SNAREs. , 1998, Science.

[24]  M. Burmeister,et al.  Mutation in AP-3 δ in the mocha Mouse Links Endosomal Transport to Storage Deficiency in Platelets, Melanosomes, and Synaptic Vesicles , 1998, Neuron.

[25]  T. Galli,et al.  A novel tetanus neurotoxin-insensitive vesicle-associated membrane protein in SNARE complexes of the apical plasma membrane of epithelial cells. , 1998, Molecular biology of the cell.

[26]  Benedikt Westermann,et al.  SNAREpins: Minimal Machinery for Membrane Fusion , 1998, Cell.

[27]  R. Scheller,et al.  Syntaxin 6 functions in trans-Golgi network vesicle trafficking. , 1997, Molecular biology of the cell.

[28]  M. Robinson,et al.  Characterization of the Adaptor-related Protein Complex, AP-3 , 1997, The Journal of cell biology.

[29]  K. Roche,et al.  AP‐3: an adaptor‐like protein complex with ubiquitous expression , 1997, The EMBO journal.

[30]  M. Gossen,et al.  Co-regulation of two gene activities by tetracycline via a bidirectional promoter. , 1995, Nucleic acids research.

[31]  R. Scheller,et al.  Two vesicle-associated membrane protein genes are differentially expressed in the rat central nervous system. , 1989, The Journal of biological chemistry.

[32]  R. Swank,et al.  Lysosomal dysfunctions associated with mutations at mouse pigment genes. , 1979, Genetics.

[33]  I. Maridonneau-Parini,et al.  The protein tyrosine kinase Hck is located on lysosomal vesicles that are physically and functionally distinct from CD63-positive lysosomes in human macrophages. , 2002, Journal of cell science.

[34]  J. Bonifacino,et al.  Altered trafficking of lysosomal proteins in Hermansky-Pudlak syndrome due to mutations in the beta 3A subunit of the AP-3 adaptor. , 1999, Molecular cell.

[35]  A. Vojtek,et al.  Ras-Raf interaction: two-hybrid analysis. , 1995, Methods in enzymology.

[36]  D. Hartley.  Cellular interactions in development : a practical approach , 1993 .

[37]  R. Swank,et al.  Platelet storage pool deficiency associated with inherited abnormalities of the inner ear in the mouse pigment mutants muted and mocha. , 1991, Blood.

[38]  D. M. Weir,et al.  Handbook of experimental immunology , 1967 .