VAMP7 controls T cell activation by regulating the recruitment and phosphorylation of vesicular Lat at TCR-activation sites

The mechanisms by which Lat (a key adaptor in the T cell antigen receptor (TCR) signaling pathway) and the TCR come together after TCR triggering are not well understood. We investigate here the role of SNARE proteins, which are part of protein complexes involved in the docking, priming and fusion of vesicles with opposing membranes, in this process. Here we found, by silencing approaches and genetically modified mice, that the vesicular SNARE VAMP7 was required for the recruitment of Lat-containing vesicles to TCR-activation sites. Our results indicated that this did not involve fusion of Lat-containing vesicles with the plasma membrane. VAMP7, which localized together with Lat on the subsynaptic vesicles, controlled the phosphorylation of Lat, formation of the TCR-Lat-signaling complex and, ultimately, activation of T cells. Our findings suggest that the transport and docking of Lat-containing vesicles with target membranes containing TCRs regulates TCR-induced signaling.

[1]  Suliana Manley,et al.  Functional nanoscale organization of signaling molecules downstream of the T cell antigen receptor. , 2011, Immunity.

[2]  T. Südhof,et al.  The synaptic vesicle cycle revisited. , 2000, Neuron.

[3]  L. Samelson,et al.  LAT The ZAP-70 Tyrosine Kinase Substrate that Links T Cell Receptor to Cellular Activation , 1998, Cell.

[4]  Lydia Danglot,et al.  Multiple roles of the vesicular‐SNARE TI‐VAMP in post‐Golgi and endosomal trafficking , 2009, FEBS letters.

[5]  C. Huynh,et al.  Identification of SNAREs Involved in Synaptotagmin VII-regulated Lysosomal Exocytosis* , 2004, Journal of Biological Chemistry.

[6]  Paige Lacy,et al.  Vesicle-associated membrane protein 7 (VAMP-7) is essential for target cell killing in a natural killer cell line. , 2008, Biochemical and biophysical research communications.

[7]  Mark M Davis,et al.  TCR and Lat are expressed on separate protein islands on T cell membranes and concatenate during activation , 2010, Nature Immunology.

[8]  Federico Gallo,et al.  Regulating secretory lysosomes. , 2006, Journal of biochemistry.

[9]  S. Singer,et al.  Polarization of the Golgi apparatus and the microtubule-organizing center within cloned natural killer cells bound to their targets. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[10]  Alexander Babich,et al.  F-actin polymerization and retrograde flow drive sustained PLCγ1 signaling during T cell activation , 2012, The Journal of cell biology.

[11]  T. Harder,et al.  Selective Accumulation of Raft-Associated Membrane Protein Lat in T Cell Receptor Signaling Assemblies , 2000, The Journal of cell biology.

[12]  T. Galli,et al.  Vesicle associated membrane protein (VAMP)‐7 and VAMP‐8, but not VAMP‐2 or VAMP‐3, are required for activation‐induced degranulation of mature human mast cells , 2008, European journal of immunology.

[13]  Olivier Lantz,et al.  Dendritic Cell Maturation Controls Adhesion, Synapse Formation, and the Duration of the Interactions with Naive T Lymphocytes , 2004, The Journal of Immunology.

[14]  Rajat Varma,et al.  T cell receptor-proximal signals are sustained in peripheral microclusters and terminated in the central supramolecular activation cluster. , 2006, Immunity.

[15]  B. Nal,et al.  Activation-induced polarized recycling targets T cell antigen receptors to the immunological synapse; involvement of SNARE complexes. , 2004, Immunity.

[16]  L. Samelson,et al.  Dynamic actin polymerization drives T cell receptor-induced spreading: a role for the signal transduction adaptor LAT. , 2001, Immunity.

[17]  I. Mérida,et al.  Diacylglycerol kinase α regulates the formation and polarisation of mature multivesicular bodies involved in the secretion of Fas ligand-containing exosomes in T lymphocytes , 2011, Cell Death and Differentiation.

[18]  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.

[19]  Helmut Grubmüller,et al.  How SNARE molecules mediate membrane fusion: recent insights from molecular simulations. , 2012, Current opinion in structural biology.

[20]  Astrid Magenau,et al.  PALM imaging and cluster analysis of protein heterogeneity at the cell surface , 2010, Journal of biophotonics.

[21]  P. Perrin,et al.  Loss of the LAT adaptor converts antigen-responsive T cells into pathogenic effectors that function independently of the T cell receptor. , 2009, Immunity.

[22]  B. Alarcón,et al.  Internalization and intracellular fate of TCR-CD3 complexes. , 2000, Critical reviews in immunology.

[23]  F. Sánchez‐Madrid,et al.  End‐binding protein 1 controls signal propagation from the T cell receptor , 2012, The EMBO journal.

[24]  T. Galli,et al.  A molecular network for the transport of the TI-VAMP/VAMP7 vesicles from cell center to periphery. , 2012, Developmental cell.

[25]  Dan R. Littman,et al.  Signal transduction by lymphocyte antigen receptors , 1994, Cell.

[26]  Astrid Magenau,et al.  Pre-existing clusters of the adaptor Lat do not participate in early T cell signaling events , 2011, Nature Immunology.

[27]  M. Waisberg,et al.  Endocytosed BCRs sequentially regulate MAPK and Akt signaling pathways from intracellular compartments , 2011, Nature Immunology.

[28]  T. Galli,et al.  [Activation-induced polarized recycling targets T cell receptors to the immunological synapse]. , 2005, Medecine sciences : M/S.

[29]  D. Olive,et al.  Dynamic recruitment of the adaptor protein LAT: LAT exists in two distinct intracellular pools and controls its own recruitment , 2004, Journal of Cell Science.

[30]  M. D'Esposito,et al.  Absence of TI-VAMP/Vamp7 Leads to Increased Anxiety in Mice , 2012, The Journal of Neuroscience.

[31]  M. Neil,et al.  Remodelling of Cortical Actin Where Lytic Granules Dock at Natural Killer Cell Immune Synapses Revealed by Super-Resolution Microscopy , 2011, PLoS biology.

[32]  J. Sibarita,et al.  TI‐VAMP/VAMP7 is required for optimal phagocytosis of opsonised particles in macrophages , 2004, The EMBO journal.

[33]  Ronald D. Vale,et al.  Single-Molecule Microscopy Reveals Plasma Membrane Microdomains Created by Protein-Protein Networks that Exclude or Trap Signaling Molecules in T Cells , 2005, Cell.

[34]  Thomas C. Südhof,et al.  The Synaptic VesicleCycle Revisited , 2000, Neuron.

[35]  C. Rudd.  Adaptors and Molecular Scaffolds in Immune Cell Signaling , 1999, Cell.

[36]  J. Husson,et al.  Force Generation upon T Cell Receptor Engagement , 2011, PloS one.

[37]  T. Galli,et al.  The Tetanus Neurotoxin‐Sensitive and Insensitive Routes to and from the Plasma Membrane: Fast and Slow Pathways? , 2005, Traffic.

[38]  F. Mollinedo,et al.  Combinatorial SNARE Complexes Modulate the Secretion of Cytoplasmic Granules in Human Neutrophils1 , 2006, The Journal of Immunology.

[39]  Mark A. A. Neil,et al.  Dynamics of Subsynaptic Vesicles and Surface Microclusters at the Immunological Synapse , 2010, Science Signaling.

[40]  V. Barr,et al.  c-Cbl-Mediated Regulation of LAT-Nucleated Signaling Complexes , 2007, Molecular and Cellular Biology.

[41]  L. Samelson,et al.  LAT palmitoylation: its essential role in membrane microdomain targeting and tyrosine phosphorylation during T cell activation. , 1998, Immunity.

[42]  L. Samelson,et al.  Association of Grb2, Gads, and phospholipase C-gamma 1 with phosphorylated LAT tyrosine residues. Effect of LAT tyrosine mutations on T cell angigen receptor-mediated signaling. , 2000, The Journal of biological chemistry.

[43]  M. Mingueneau,et al.  Lymphoproliferative disorders involving T helper effector cells with defective LAT signalosomes , 2010, Seminars in Immunopathology.

[44]  F. Miró,et al.  Cytokine Secretion by CD4+ T Cells at the Immunological Synapse Requires Cdc42-Dependent Local Actin Remodeling but Not Microtubule Organizing Center Polarity , 2012, The Journal of Immunology.

[45]  T. Svitkina,et al.  Natural Killer Cell Lytic Granule Secretion Occurs through a Pervasive Actin Network at the Immune Synapse , 2011, PLoS biology.

[46]  Michael Loran Dustin,et al.  Essential role of ubiquitin and TSG101 protein in formation and function of the central supramolecular activation cluster. , 2010, Immunity.

[47]  C. Hivroz,et al.  In the immune synapse, ZAP-70 controls T cell polarization and recruitment of signaling proteins but not formation of the synaptic pattern. , 2002, Immunity.

[48]  Eric O Long,et al.  Essential role of LAT in T cell development. , 1999, Immunity.

[49]  T. Südhof,et al.  Cellubrevin is a ubiquitous tetanus-toxin substrate homologous to a putative synaptic vesicle fusion protein , 1993, Nature.

[50]  M. A. Alonso,et al.  MAL Protein Controls Protein Sorting at the Supramolecular Activation Cluster of Human T Lymphocytes , 2011, The Journal of Immunology.

[51]  M. Dahan,et al.  Role of TI-VAMP and CD82 in EGFR cell-surface dynamics and signaling , 2010, Development.