Control of vesicular trafficking

Control of vesicular trafficking by synaptic proteins in neuronal and neuroendocrine cells,
its changes during pathology

In the frame of INTAS project

# INTAS 01 2095

Prof. E. Neher (Co-ordinator) Department of membrane Biophysics Max-Planck-Institut fur Biophysikalische Chemie, Goettingen, Germany Prof. R. Burgyone Department of Physiology University of Liverpool, United Kingdom Prof. R. Jahn Department of Neurobiology, Max-Planck-Institut fur Biophysikalische Chemie, Goettingen, Germany Prof. N. Brose Department of Molecular Neurobiology Max-Planck-Institute for Experimental Medicine, Goettingen, Germany

Prof. P. Kostyuk Bogomoletz Institute of Physiology, Kiev, Ukraine Dr. E. Lukyanetz International Center for Molecular Physiology, Kiev, Ukraine Prof. Volkov G. Palladin Institute of Biochemistry, Kiev, Ukraine

The goal of the proposed project is the investigation of the role of synaptic proteins in disposition and trafficking of secretory vesicles in synapses of central neurons and neurosecretory cells. Recent investigations including the work of the project’s participants have shown that vesicular transport events in neuroendocrine cells and presynaptic terminals are mediated by families of specialized membrane proteins in the vesicle (v-SNAREs) and in the target cytoplasmic membrane (t-SNAREs). A variety of such proteins has been already described in distinct preparations; however, their precise localization and their role in vesicular trafficking during functional changes of the cells remain ambiguous. Therefore, in planned experiments, it is aimed to investigate the role of synaptic proteins in trafficking of secretory vesicles by means of core SNARE proteins - vesicular (synaptobrevin/VAMPs) and plasma membrane (syntaxins/SNAP-25) synaptic proteins, heterotrimeric G-proteins as well as novel presynaptic proteins (Munc13, Munc-18, CAPS, Csp, Doc2 and complexins). Both neuronal and neuroendocrine cells will be used in the planned experiments; comparison of these systems in terms of synaptic proteins’ specificity to different types of secretory vesicles has not been previously discussed. It is expected that the results of the project will help us to characterize behavior and specificity of the synaptic proteins involved in the process of trafficking of different types of vesicles in order to understand basic principles of neurotransmission, and their changes during pathology.

Meetings in the frame of the project

File:Control of vesicular trafficking.pdf

Kiev INTAS Meeting, 2004

List of publications of Teams on the topic of the project

1. Antonin W., Dulubova, I., Arac, D., Pabst, S., Plitzner, J., Rizo, J., Jahn, R. (2002) The N-terminal domains of syntaxin 7 and vti1b form three-helix bundles that differ in their ability to regulate SNARE complex assembly. J. Biol. Chem. 277, 36449-36456 # Antonin, W., Fasshauer, D., Becker, S., Jahn, R., Schneider, T.R.. (2002) Crystal structure of the endosomal SNARE complex reveals common structural principles of all SNAREs. Nature Struct. Biol., 9, 107-111
2. Antonin, W., Wagner, M., Riedel, D., Brose, N., Jahn, R. (2002) Loss of the zymogen granule protein syncollin affects pancreatic protein synthesis and transport but not secretion. Mol. Cell. Biol. 22, 1545-1554
3. Archer, D.A., M.E. Graham, and R.D. Burgoyne. 2002. Complexin regulates the closure of the fusion pore during regulated exocytosis. J. Biol.Chem. 277:18249-18252.
4. Bacia, K., Schuette, C.G., Kahya, N., Jahn, R., Schwille, P. (2004) SNAREs prefer liqued-disordered over "raft"-(liquid ordered) domains when reconstituted into giant unilamellar vesicles. J. Biol. Chem., in press
5. Barclay, J.W., T.J. Craig, R.J. Fisher, L.F. Ciufo, G.J.O. Evans, A. Morgan, and R.D. Burgoyne. (2003) Phosphorylation of Munc18 by protein kinase C regulates the kinetics of exocytosis. J. Biol.Chem. 278:10538-10545.
6. Brose, N. and E. Neher (2002). Specificity emerges in the dissection of diacylglyerol- and protein kinase C-mediated signalling pathways. PNAS 99, 16522-16523.
7. Brose, N. and Rosenmund, C. (2002) Move over protein kinase C, you've got company: alternative cellular effectors of diacylglycerol and phorbol esters. J. Cell Sci. 115, 4399-4411.
8. Brose, N., Betz, A. and Wegmeyer, H. (2004) Divergent and convergent signaling by the diacylglycerol second messenger pathway in mammals. Curr. Op. Neurobiol. 14, 328-340.
9. Bruns, D., Jahn, R.: Molecular determinants of exocytosis (2002) Pfl?gers Arch. - Eur. J. Physiol, 443, 333-338
10. Burgoyne, R.D., and A. Morgan. (2003) Secretory granule exocytosis. Physiol. Rev. 83:581-632.
11. Chandra, S., Chen, X., Rizo, J., Jahn, R., S?dhof, T.C. (2003) A broken -helix in folded -synuclein. J. Biol. Chem. 278, 15313-15318
12. Craig, T.J., Evans, G.J.O. and Morgan, A. (2003) Physiological regulation of Munc18/nSec1 phosphorylation on serine-313 J. Neurochem. 86, 1450-1457.
13. Evans, G.J.O., M.C. Wilkinson, M.E. Graham, K.M. Turner, L.H. Chamberlain, R.D. Burgoyne, and A. Morgan (2001) Phosphorylation of cysteine string protein by PKA: implications for the modulation of exocytosis. J. Biol. Chem. 276:47877-47885.
14. Fasshauer, D., Antonin, W., Subramaniam, V., Jahn, R. (2002) SNARE assembly and disassembly exhibit a pronounced hysteresis (2002) Nature Struct. Biol. 9, 144-151
15. Fasshauer, D., Sch?tte, C., Jahn, R. (2001) Mechanisms of membrane fusion. B.I.F. Futura 16, 13-23
16. Fisher, R.J., J. Pevsner, and R.D. Burgoyne (2001) Control of fusion pore dynamics during exocytosis by Munc18. Science. 291:875-878.
17. Gallwitz, D., Jahn, R. (2003) The riddle of the Sec1/Munc-18 proteins - new twists added to their interactions with SNAREs. Trends Biochem. Sci. 28, 113-116
18. Graham, M.E., Barclay, J.W. & Burgoyne, R.D. (2004) Syntaxin/Munc18 interactions in the late events during vesicle fusion and release in exocytosis. J. Biol. Chem. In press.
19. Graham, M.E., D.W. O'Callaghan, H.T. McMahon, and R.D. Burgoyne (2002)Dynamin-dependent and dynamin-independent processes contribute to the regulation of single vesicle release kinetics and quantal size. Proc. Natl. Acad. Sci. USA. 99:7124-7129.
20. Hatsuzawa, K., Lang, T., Fasshauer, D., Bruns, D., Jahn, R. (2003) The R-SNARE motif of tomosyn forms SNARE core complexes with syntaxin 1 and SNAP-25 and down-regulates exocytosis. J. Biol. Chem. 278, 31159-31166
21. Holroyd, P., Lang, T., Wenzel, D., De Camilli, P., Jahn, R. (2002) Imaging direct, dynamin-dependent recapture of fusing secretory granules on plasma membrane lawns from PC12 cells. Proc. Natl. Acad. Sci. USA 99, 16806-16811
22. Holt, M., Jahn, R. (2004) Synaptic vesicles in the fast lane. Science 303, 1986-1987
23. Hopt., A., Korte, S., Fink, H., Panne, U., Niessner, R., Jahn, R., Kretzschmar, H., Herms, J. (2003) Methods for studying synaptosomal copper release. J. Neurosci. Meth. 128, 159-172
24. Jahn, R. (2004) Principles of membrane fusion and exocytosis. Ann. N.Y.Acad. Sci. 1014, 170-178
25. Jahn, R., Grubm?ller, H. (2002) Membrane fusion. Curr. Opinion in Cell Biology, 14, 488-495 (2002)
26. Jahn, R., Lang, T., S?dhof, T.C. (2003) Membrane fusion. Cell 112, 519-533
27. Jahn, R., Schuman, E. (2001) Neuronal and glial cell biology. Curr. Opinion in Neurobiology 11, 533-535.
28. Junge, H., Rhee, J.-S., Jahn, O., Varoqueaux, F., Spiess, J., Waxham, M.N., Rosenmund, C. and Brose, N. (2004) Calmodulin and Munc13 form a Ca2+-sensor/effector complex that controls short-term synaptic plasticity. Cell, in press.
29. Khvotchev, M.V., Ren, M., Takamori, S., Jahn, R., S?dhof, T.C. (2003) Divergent functions of neuronal Rab11b in Ca2+-regulated vs. constitutive exocytosis. J. Neurosci. 23, 10531-10539
30. Koval L.M., Yavorskaya E.N., Lukyanetz E.A. 2001. Electron microscopic evidence for multiple types of secretory vesicles in bovine chromaffin cells. General. Compar. Endocrynol. 121, (3), 261-277.
31. Lang, T., Bruns, D., Wenzel, D., Riedel, D., Holroyd, P., Thiele, C., Jahn, R. (2001) SNAREs are concentrated in cholesterol-dependent clusters that define docking and fusion sites for exocytosis. EMBO J. 20, 2202-2213
32. Lang, T., Jahn, R. (2002) Fusion of vacuoles - where are the membranes, and where are the holes? (Preview) Dev. Cell 2, 257-259
33. Lang, T., Margittai, M., H?lzler, H., Jahn, R. (2002) SNAREs in native plasma membranes are active and readily form core complexes with endogenous and exogenous SNAREs. J. Cell Biol. 158, 751-760
34. Lukyanetz E. (2002) Vesicular mechanisms of calcium-dependent exocytosis of neurotransmitters. Arch. Clini. Exper. Medicine. v. 11, № 1, 2002, p. 10-14.
35. Lukyanetz E.A. 2001. Intracellular events during depolarization-induced exocytosis in chromaffin cells. In: Calcium Signalling, Morad M., Kostyuk P. eds. NATO Science Series, Series I: Life and Behavioural Sciences. V.331, IOS Press Ohmsha, Amsterdam, The Netherlands. p. 174-182.
36. Lukyanetz E.A. 2001. Intracellular mechanism of calcium-dependent neurotransmitter release. Bogomoetz-Nencki Meeting, Sulejow, 2001, p. L4.
37. Lukyanetz E.A., Pochynyuk O.M., Zaika O.L., 2002. Amperometric evidence about participation of several types of vesicles in exocytosis from chromaffin cells. J. Physiology (Paris), 96, p.147-148.
38. Lukyanetz E.A., Stanika R.I., Koval L.M., Yavorskaya E.N., Kostyuk P.G. 2004. Hypoxia-induced intracellular calcium oscillations in rat chromaffin cells. Br.Res, in press
39. Lukyanetz E.A., Zaika O.L., Pochynyuk O.M., Kostyuk P.G. 2001. Two classes of vesicles participate in induced exocytosis from isolated rat chromaffin cells. J. Physiol. 231P, 162P
40. Lukyanetz E.A.. Different secretory vesicles can be involved in depolarization-evoked exocytosis. 2001. Biochem.Biophys.Res.Com. 288 (4), p.844-848.
41. Lukyanetz, EA, Pochynyuk, OM, Zaika OM. 2001. Evidence for participation of different types of vesicles in stimulus-induced exocytosis in chromaffin cells. XXXIV International Congress of Physiological Sciences, New Zeland, August 26-31, 2001, IUPS
42. Margittai, M., Fasshauer, D., Jahn, R., Langen, R. (2003) Habc domain and SNARE core complex are connected by a highly flexible linker. Biochemistry, 42, 4009-4014
43. Margittai, M., Fasshauer, D., Pabst, S., Jahn, R., Langen, R. (2001) Homo- and heterooligomeric SNARE complexes studied by site-directed spin labeling. J. Biol. Chem. 276, 13169-13177
44. Margittai, M., Widengren, J., Schweinberger, E., Schr?der, G.F., Fasshauer, D., Felekyan, S., Haustein, E., K?nig, M., Grubm?ller, H., Jahn, R., Seidel, C.A.M. (2003) Single-molecule FRET reveals a dynamic equilibirum between closed and open conformations of syntaxin-1. Proc. Natl. Acad. Sci. USA 100, 15516-15521
45. Milosevic, I., J.B. S?rensen, T. Lang, M. Krauss, G. Nagy, V. Haucke, R. Jahn and E. Neher (2004). Plasmalemmal PI(4,5)P2 level regulates the releasable vesicle pool size in chromaffin cells. (submitted)
46. Mueller V.J., Wienisch M., Nehring R.B., Klingauf J. (2004). Monitoring clathrin-mediated endocytosis during synaptic activity . J. Neurosci. 24(8), 2004-12.
47. Nagy, G., Reim, K., Matti, U., Brose, N., Binz, T., Rettig, J., Neher, E. and J.B. S?rensen (2003). Regulation of releasable vesicle pool sizes by protein kinase A-dependent phosphorylation of SNAP-25. Neuron 41, 417-429.
48. Nagy, G., U. Matti, R.B. Nehring, T. Binz, J. Rettig, E. Neher and J.B. Soerensen (2002). Protein kinase C-dependent phosphorylation of synaptosome-associated protein of 25 kDa at Ser187 potentiates vesicle recruitment. J. Neurosci. 22, 9278-9286.
49. Pabst, S., Margittai, M., Vainius, D., Langen, R., Jahn, R., Fasshauer, D. (2002) Rapid and selective binding to the synaptic SNARE complex suggests a modulatory role of complexins in neuroexocytosis. J. Biol. Chem., 277, 7838-7848
50. Pochynyuk O., Zaika O., Lukyanetz E. (2002) The role of mitochondria in generation of acetylcholine-induced calcium transients in rat chromaffin cells. Neurophysiology, 34, No 2/3, 2002, p. 217-219.
51. Pochynyuk O.M., Zaika O.L, Bidnenko V.M., Kostyuk P.G., Lukyanetz E.A. 2004. Difference in exocytotic responses in rat chromaffin cells during different types of stimulation. In press.
52. Pochynyuk O.M., Zaika O.L, Kostyuk P.G., Lukyanetz E.A. 2004.
53. Pochynyuk, OM, Zaika OM, Lukyanetz, EA. Amperometrical studies of secretory responses of rat chromaffin cells induced by acetylcholine receptor agonists. 2001, Arch.Сlin.Exp.Medicine. 10 (2), 206.
54. Rettig, J. and E. Neher (2002). Emerging roles of presynaptic proteins in Ca++-triggered exocytosis. Science 298, 781-785.
55. Rhee, J.-S., Betz, A., Pyott, S., Reim, K., Varoqueaux, F., Augustin, I., Hesse, D., S?dhof, T.C., Takahashi, M., Rosenmund, C. and Brose, N. (2002) Phorbol ester- and diacylglycerol-induced augmentation of transmitter release is mediated by Munc13s and not by PKCs. Cell 108, 121-133.
56. Riedel, D., Antonin, W., Fernandez-Chacon, R., Jo, T., Geppert, M., Valentjin, J.A., Valentjin, K., Jamieson, J.D., S?dhof, T.C., Jahn, R. (2002) Rab3D is not required for exocrine exocytosis, but maintenance of normally sized secretory granules. Mol. Cell. Biol. 22, 6487-6497
57. Rosenmund, C., Sigler, A., Augustin, I., Reim, K., Brose, N. and Rhee, J.-S. (2002) Differential control of vesicle priming and short-term plasticity by Munc13 isoforms. Neuron 33, 411-424.
58. Ro?ner, S., Fuchsbrunner, K., Lange-Dohna, C., Hartlage-R?bsamen, M., Bigl, V., Betz, A., Reim, K. and Brose, N. (2004) Munc13-1-mediated vesicle priming contributes to secretory APP processing. J. Biol. Chem. 279, 27841-27844.
59. Schl?ter, O.M., Khovtchev, M., Jahn, R., S?dhof, T.C. (2002) Localization versus function of Rab3 proteins. Evidence for a common regulatory role in controlling fusion. J. Biol. Chem. 277, 40919-40929
60. Schl?ter, O.M., Schmitz, F., Jahn, R., Rosenmund, C., S?dhof, T.C.: A complete genetic analysis of neuronal Rab3 function. J. Neurosci., in press.
61. Schuette, C.G., Hatsuzawa, K., Margittai, M., Stein, A., Riedel, D., K?ster, P., K?nig, M., Seidel, C.A.M., Jahn, R. (2004) Determinants of liposome fusion mediated by synaptic SNARE proteins. Proc. Natl. Acad. Sci. 101, 2858-2863
62. Sheu, L., Pasyk, E.A., Ji, J., Huang, X., Gao, X., Varoqueaux, F., Brose, N. and Gaisano, H.Y. (2003) Regulation of insulin exocytosis by Munc13-1. J. Biol. Chem. 278, 27556-27563.
63. S?rensen, J., Nagy, G., Varoqueaux, F., Nehring, R.B., Brose, N., Wilson, M.C. and E. Neher (2003). Differential control of the releasable vesicle pools by SNAP-25 splice variants and SNAP-23. Cell 114, 75-86.
64. S?rensen, J.B., Matti, U., Wei, S.-H., Nehring, R.B., Voets, T., Ashery, U., Binz, T., Neher, E., and Rettig, J. (2002). The SNARE protein SNAP-25 is linked to fast calcium triggering of exocytosis. PNAS 99, 1627-1632.
65. Speidel, D., Varoqueaux, F., Enk, C., Nojiri, M., Grishanin, R.N., Martin, T.F., Hofmann, K., Brose, N. and Reim, K. (2003) A family of Ca2+-dependent activator proteins for secretion: Comparative analysis of structure, expression, localization, and function. J. Biol. Chem. 278, 52802-52809.
66. Stobrawa, S.M., Breiderhoff, T., Takamori,S., Engel,D., Schweizer, M., Zdebik, A.A., B?sl, M.R., Ruether, K., Jahn, H., Draguhn, R., Jahn, R., Jentsch, T.J. (2001) Disruption of ClC-3, a chloride channel expressed on synaptic vesicles, leads to a loss of the hippocampus. Neuron 29, 185-196
67. Takamori, S., Rhee, J.S., Rosenmund, C., Jahn, R.: (2001) Identification of Differentiation-Associated Brain-Specific Phosphate Transporter as a Second Vesicular Glutamate Transporter (VGLUT2). J. Neurosci. 21:RC 182 (1-6)
68. Takamori, S.,Malherbe, P., Broger, C., Jahn, R. (2002) Molecular cloning and functional characterization of human vesicular glutamate transporter 3. EMBO Reports 3, 798-803
69. Tokar S.L., Koval L.M., Yavorskaya E.N., Lukyanetz E.A. Calcium-induced ultrastructural interactions in adrenocortical cells. 2001, Arch.Сlin.Exp.Medicine. 10 (2), 224.
70. Varoqueaux, F., Sigler, A., Rhee, J.-S., Brose, N., Enk, C., Reim, K. and Rosenmund, C. (2002) Total arrest of spontaneous and evoked synaptic transmission but normal synaptogenesis in the absence of Munc13-mediated vesicle priming. Proc. Natl. Acad. Sci. U.S.A. 99, 9037-9042.
71. Vites, O., Rhee, J.-S., Schwarz, M., Rosenmund, C., Jahn, R.: Re-investigation of the role of snapin in neurotransmitter release. J. Biol. Chem., in press
72. Wojcik, S.M., Rhe, J.S., Herzog, E., Sigler, A., Jahn, R., Takamori, S., Brose, N., Rosenmund, C. (2004) An essential role for VGLUT1 in postnatal development and control of quantal size. Proc. Natl. Acad. Sci. USA 101, 58-63.
73. Zaika O., Pochynyuk O., Lukyanetz E. (2002) Comparative studies of calcium transients induced by acetylcholine in rat chromaffin cells. Neurophysiology, 34, No 2/3, 2002, p. 270-272.
74. Zaika O.L., Pochynyuk O.M. Lukyanetz E.A. 2001. Electrochemical studies of induced secretion of catecholamines from single vesicles of rat chromaffin cells. Ukr. Biochem J. Vol.73(4) pp.69-72
75. Zaika O.L., Pochynyuk O.M., Lukyanetz E.A. 2002. Investigation of calcium transients induced by acetylcholine in isolated chromaffin cells of rat. Fiziol Zh., 48(2), p.5-6.
76. Zaika O.L., Pochynyuk O.M., Lukyanetz E.A. Altering of kinetics of neurotransmitter release measured by amperometry during different types of stimulation in rat chromaffin cells. Bogomoletz-Nencki Meeting, Sulejow, 2001, P23.
77. Zhang, H., K.W. Peters, F. Sun, C.R. Marino, J. Lang, R.D. Burgoyne, and R.A. Frizell (2002) Cysteine string protein interacts with and modulates the maturation of CFTR. J. Biol.Chem. 277:28948-28958.