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EMBO J. Jan 2, 1998; 17(1): 127–139.
PMCID: PMC1170364

ROP, the Drosophila Sec1 homolog, interacts with syntaxin and regulates neurotransmitter release in a dosage-dependent manner.

Abstract

The Sec1 family of proteins is thought to function in both non-neuronal and neuronal secretion, although the precise role of this protein family has not been defined. Here, we study the function of ROP, the Drosophila Sec1 homolog, in neurotransmitter release. Electrophysiological analyses of transgenic lines overexpressing ROP and syntaxin, a presynaptic membrane protein, indicate that ROP interacts with syntaxin in vivo. Characterization of four point mutations in ROP shows that they fall into two phenotypic classes. Two mutations cause a dramatic reduction in both evoked and spontaneous neurotransmitter release. In contrast, the other two mutations reveal an increase in evoked neurotransmission. Our data further show that neurotransmission is highly sensitive to the levels of ROP function. Studies on heterozygote animals indicate that half the amount of wild-type ROP results in a dramatic decrease in evoked and spontaneous exocytosis. Taken together, these results suggest that ROP interacts with syntaxin in vivo and is a rate-limiting regulator of exocytosis that performs both positive and inhibitory functions in neurotransmission.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aalto MK, Ronne H, Keränen S. Yeast syntaxins Sso1p and Sso2p belong to a family of related membrane proteins that function in vesicular transport. EMBO J. 1993 Nov;12(11):4095–4104. [PMC free article] [PubMed]
  • Bellen HJ, O'Kane CJ, Wilson C, Grossniklaus U, Pearson RK, Gehring WJ. P-element-mediated enhancer detection: a versatile method to study development in Drosophila. Genes Dev. 1989 Sep;3(9):1288–1300. [PubMed]
  • Ben-Ari Y, Aniksztejn L, Bregestovski P. Protein kinase C modulation of NMDA currents: an important link for LTP induction. Trends Neurosci. 1992 Sep;15(9):333–339. [PubMed]
  • Bennett MK, Scheller RH. The molecular machinery for secretion is conserved from yeast to neurons. Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):2559–2563. [PMC free article] [PubMed]
  • Bennett MK, Calakos N, Scheller RH. Syntaxin: a synaptic protein implicated in docking of synaptic vesicles at presynaptic active zones. Science. 1992 Jul 10;257(5067):255–259. [PubMed]
  • Bezprozvanny I, Scheller RH, Tsien RW. Functional impact of syntaxin on gating of N-type and Q-type calcium channels. Nature. 1995 Dec 7;378(6557):623–626. [PubMed]
  • Bittner MA, Bennett MK, Holz RW. Evidence that syntaxin 1A is involved in storage in the secretory pathway. J Biol Chem. 1996 May 10;271(19):11214–11221. [PubMed]
  • Blasi J, Chapman ER, Link E, Binz T, Yamasaki S, De Camilli P, Südhof TC, Niemann H, Jahn R. Botulinum neurotoxin A selectively cleaves the synaptic protein SNAP-25. Nature. 1993 Sep 9;365(6442):160–163. [PubMed]
  • Blasi J, Chapman ER, Yamasaki S, Binz T, Niemann H, Jahn R. Botulinum neurotoxin C1 blocks neurotransmitter release by means of cleaving HPC-1/syntaxin. EMBO J. 1993 Dec;12(12):4821–4828. [PMC free article] [PubMed]
  • Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. [PMC free article] [PubMed]
  • Broadie KS, Bate M. Development of the embryonic neuromuscular synapse of Drosophila melanogaster. J Neurosci. 1993 Jan;13(1):144–166. [PubMed]
  • Broadie K, Prokop A, Bellen HJ, O'Kane CJ, Schulze KL, Sweeney ST. Syntaxin and synaptobrevin function downstream of vesicle docking in Drosophila. Neuron. 1995 Sep;15(3):663–673. [PubMed]
  • Chapman ER, An S, Barton N, Jahn R. SNAP-25, a t-SNARE which binds to both syntaxin and synaptobrevin via domains that may form coiled coils. J Biol Chem. 1994 Nov 4;269(44):27427–27432. [PubMed]
  • Dascher C, Ossig R, Gallwitz D, Schmitt HD. Identification and structure of four yeast genes (SLY) that are able to suppress the functional loss of YPT1, a member of the RAS superfamily. Mol Cell Biol. 1991 Feb;11(2):872–885. [PMC free article] [PubMed]
  • Dascher C, Matteson J, Balch WE. Syntaxin 5 regulates endoplasmic reticulum to Golgi transport. J Biol Chem. 1994 Nov 25;269(47):29363–29366. [PubMed]
  • DiAntonio A, Schwarz TL. The effect on synaptic physiology of synaptotagmin mutations in Drosophila. Neuron. 1994 Apr;12(4):909–920. [PubMed]
  • Ferro-Novick S, Jahn R. Vesicle fusion from yeast to man. Nature. 1994 Jul 21;370(6486):191–193. [PubMed]
  • Fujita Y, Sasaki T, Fukui K, Kotani H, Kimura T, Hata Y, Südhof TC, Scheller RH, Takai Y. Phosphorylation of Munc-18/n-Sec1/rbSec1 by protein kinase C: its implication in regulating the interaction of Munc-18/n-Sec1/rbSec1 with syntaxin. J Biol Chem. 1996 Mar 29;271(13):7265–7268. [PubMed]
  • Galey FR, Nilsson SE. A new method for transferring sections from the liquid surface of the trough through staining solutions to the supporting film of a grid. J Ultrastruct Res. 1966 Feb;14(3):405–410. [PubMed]
  • Ganetzky B, Wu CF. Neurogenetic analysis of potassium currents in Drosophila: synergistic effects on neuromuscular transmission in double mutants. J Neurogenet. 1983 Sep;1(1):17–28. [PubMed]
  • Garcia EP, Gatti E, Butler M, Burton J, De Camilli P. A rat brain Sec1 homologue related to Rop and UNC18 interacts with syntaxin. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2003–2007. [PMC free article] [PubMed]
  • Garcia EP, McPherson PS, Chilcote TJ, Takei K, De Camilli P. rbSec1A and B colocalize with syntaxin 1 and SNAP-25 throughout the axon, but are not in a stable complex with syntaxin. J Cell Biol. 1995 Apr;129(1):105–120. [PMC free article] [PubMed]
  • Halachmi N, Feldman M, Kimchie Z, Lev Z. Rop and Ras2, members of the Sec1 and Ras families, are localized in the outer membranes of labyrinthine channels and vesicles of Drosophila nephrocyte, the Garland cell. Eur J Cell Biol. 1995 Jul;67(3):275–283. [PubMed]
  • Hanson PI, Otto H, Barton N, Jahn R. The N-ethylmaleimide-sensitive fusion protein and alpha-SNAP induce a conformational change in syntaxin. J Biol Chem. 1995 Jul 14;270(28):16955–16961. [PubMed]
  • Harrison SD, Broadie K, van de Goor J, Rubin GM. Mutations in the Drosophila Rop gene suggest a function in general secretion and synaptic transmission. Neuron. 1994 Sep;13(3):555–566. [PubMed]
  • Hata Y, Slaughter CA, Südhof TC. Synaptic vesicle fusion complex contains unc-18 homologue bound to syntaxin. Nature. 1993 Nov 25;366(6453):347–351. [PubMed]
  • Hawkins RD, Kandel ER, Siegelbaum SA. Learning to modulate transmitter release: themes and variations in synaptic plasticity. Annu Rev Neurosci. 1993;16:625–665. [PubMed]
  • Hayashi T, McMahon H, Yamasaki S, Binz T, Hata Y, Südhof TC, Niemann H. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly. EMBO J. 1994 Nov 1;13(21):5051–5061. [PMC free article] [PubMed]
  • Hosono R, Hekimi S, Kamiya Y, Sassa T, Murakami S, Nishiwaki K, Miwa J, Taketo A, Kodaira KI. The unc-18 gene encodes a novel protein affecting the kinetics of acetylcholine metabolism in the nematode Caenorhabditis elegans. J Neurochem. 1992 Apr;58(4):1517–1525. [PubMed]
  • Hunt JM, Bommert K, Charlton MP, Kistner A, Habermann E, Augustine GJ, Betz H. A post-docking role for synaptobrevin in synaptic vesicle fusion. Neuron. 1994 Jun;12(6):1269–1279. [PubMed]
  • Jahn R, Südhof TC. Synaptic vesicles and exocytosis. Annu Rev Neurosci. 1994;17:219–246. [PubMed]
  • Jan LY, Jan YN. Properties of the larval neuromuscular junction in Drosophila melanogaster. J Physiol. 1976 Oct;262(1):189–214. [PMC free article] [PubMed]
  • Jan YN, Jan LY, Dennis MJ. Two mutations of synaptic transmission in Drosophila. Proc R Soc Lond B Biol Sci. 1977 Jul 28;198(1130):87–108. [PubMed]
  • Kandel ER. Calcium and the control of synaptic strength by learning. Nature. 1981 Oct 29;293(5835):697–700. [PubMed]
  • Larkman AU, Jack JJ. Synaptic plasticity: hippocampal LTP. Curr Opin Neurobiol. 1995 Jun;5(3):324–334. [PubMed]
  • Littleton JT, Bellen HJ, Perin MS. Expression of synaptotagmin in Drosophila reveals transport and localization of synaptic vesicles to the synapse. Development. 1993 Aug;118(4):1077–1088. [PubMed]
  • Littleton JT, Stern M, Schulze K, Perin M, Bellen HJ. Mutational analysis of Drosophila synaptotagmin demonstrates its essential role in Ca(2+)-activated neurotransmitter release. Cell. 1993 Sep 24;74(6):1125–1134. [PubMed]
  • McMahon HT, Missler M, Li C, Südhof TC. Complexins: cytosolic proteins that regulate SNAP receptor function. Cell. 1995 Oct 6;83(1):111–119. [PubMed]
  • Meffert MK, Calakos NC, Scheller RH, Schulman H. Nitric oxide modulates synaptic vesicle docking fusion reactions. Neuron. 1996 Jun;16(6):1229–1236. [PubMed]
  • Nagamatsu S, Fujiwara T, Nakamichi Y, Watanabe T, Katahira H, Sawa H, Akagawa K. Expression and functional role of syntaxin 1/HPC-1 in pancreatic beta cells. Syntaxin 1A, but not 1B, plays a negative role in regulatory insulin release pathway. J Biol Chem. 1996 Jan 12;271(2):1160–1165. [PubMed]
  • Novick P, Field C, Schekman R. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell. 1980 Aug;21(1):205–215. [PubMed]
  • Novick P, Brennwald P, Walworth NC, Kabcenell AK, Garrett M, Moya M, Roberts D, Müller H, Govindan B, Bowser R. The cycle of SEC4 function in vesicular transport. Ciba Found Symp. 1993;176:218–232. [PubMed]
  • Orita S, Sasaki T, Naito A, Komuro R, Ohtsuka T, Maeda M, Suzuki H, Igarashi H, Takai Y. Doc2: a novel brain protein having two repeated C2-like domains. Biochem Biophys Res Commun. 1995 Jan 17;206(2):439–448. [PubMed]
  • Oyler GA, Higgins GA, Hart RA, Battenberg E, Billingsley M, Bloom FE, Wilson MC. The identification of a novel synaptosomal-associated protein, SNAP-25, differentially expressed by neuronal subpopulations. J Cell Biol. 1989 Dec;109(6 Pt 1):3039–3052. [PMC free article] [PubMed]
  • Pevsner J, Hsu SC, Braun JE, Calakos N, Ting AE, Bennett MK, Scheller RH. Specificity and regulation of a synaptic vesicle docking complex. Neuron. 1994 Aug;13(2):353–361. [PubMed]
  • Pevsner J, Hsu SC, Scheller RH. n-Sec1: a neural-specific syntaxin-binding protein. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1445–1449. [PMC free article] [PubMed]
  • Protopopov V, Govindan B, Novick P, Gerst JE. Homologs of the synaptobrevin/VAMP family of synaptic vesicle proteins function on the late secretory pathway in S. cerevisiae. Cell. 1993 Sep 10;74(5):855–861. [PubMed]
  • Risinger C, Blomqvist AG, Lundell I, Lambertsson A, Nässel D, Pieribone VA, Brodin L, Larhammar D. Evolutionary conservation of synaptosome-associated protein 25 kDa (SNAP-25) shown by Drosophila and Torpedo cDNA clones. J Biol Chem. 1993 Nov 15;268(32):24408–24414. [PubMed]
  • Salminen A, Novick PJ. A ras-like protein is required for a post-Golgi event in yeast secretion. Cell. 1987 May 22;49(4):527–538. [PubMed]
  • Salzberg A, Cohen N, Halachmi N, Kimchie Z, Lev Z. The Drosophila Ras2 and Rop gene pair: a dual homology with a yeast Ras-like gene and a suppressor of its loss-of-function phenotype. Development. 1993 Apr;117(4):1309–1319. [PubMed]
  • Sassa T, Ogawa H, Kimoto M, Hosono R. The synaptic protein UNC-18 is phosphorylated by protein kinase C. Neurochem Int. 1996 Nov;29(5):543–552. [PubMed]
  • Schiavo G, Benfenati F, Poulain B, Rossetto O, Polverino de Laureto P, DasGupta BR, Montecucco C. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature. 1992 Oct 29;359(6398):832–835. [PubMed]
  • Schulze KL, Bellen HJ. Drosophila syntaxin is required for cell viability and may function in membrane formation and stabilization. Genetics. 1996 Dec;144(4):1713–1724. [PMC free article] [PubMed]
  • Schulze KL, Littleton JT, Salzberg A, Halachmi N, Stern M, Lev Z, Bellen HJ. rop, a Drosophila homolog of yeast Sec1 and vertebrate n-Sec1/Munc-18 proteins, is a negative regulator of neurotransmitter release in vivo. Neuron. 1994 Nov;13(5):1099–1108. [PubMed]
  • Schulze KL, Broadie K, Perin MS, Bellen HJ. Genetic and electrophysiological studies of Drosophila syntaxin-1A demonstrate its role in nonneuronal secretion and neurotransmission. Cell. 1995 Jan 27;80(2):311–320. [PubMed]
  • Schuster CM, Davis GW, Fetter RD, Goodman CS. Genetic dissection of structural and functional components of synaptic plasticity. I. Fasciclin II controls synaptic stabilization and growth. Neuron. 1996 Oct;17(4):641–654. [PubMed]
  • Schwartz JH. Cognitive kinases. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8310–8313. [PMC free article] [PubMed]
  • Sesack SR, Snyder CL. Cellular and subcellular localization of syntaxin-like immunoreactivity in the rat striatum and cortex. Neuroscience. 1995 Aug;67(4):993–1007. [PubMed]
  • Sheng ZH, Rettig J, Cook T, Catterall WA. Calcium-dependent interaction of N-type calcium channels with the synaptic core complex. Nature. 1996 Feb 1;379(6564):451–454. [PubMed]
  • Söllner T, Whiteheart SW, Brunner M, Erdjument-Bromage H, Geromanos S, Tempst P, Rothman JE. SNAP receptors implicated in vesicle targeting and fusion. Nature. 1993 Mar 25;362(6418):318–324. [PubMed]
  • Stewart BA, Atwood HL, Renger JJ, Wang J, Wu CF. Improved stability of Drosophila larval neuromuscular preparations in haemolymph-like physiological solutions. J Comp Physiol A. 1994 Aug;175(2):179–191. [PubMed]
  • Südhof TC. The synaptic vesicle cycle: a cascade of protein-protein interactions. Nature. 1995 Jun 22;375(6533):645–653. [PubMed]
  • Sweeney ST, Broadie K, Keane J, Niemann H, O'Kane CJ. Targeted expression of tetanus toxin light chain in Drosophila specifically eliminates synaptic transmission and causes behavioral defects. Neuron. 1995 Feb;14(2):341–351. [PubMed]
  • Trimble WS, Cowan DM, Scheller RH. VAMP-1: a synaptic vesicle-associated integral membrane protein. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4538–4542. [PMC free article] [PubMed]
  • Verhage M, de Vries KJ, Røshol H, Burbach JP, Gispen WH, Südhof TC. DOC2 proteins in rat brain: complementary distribution and proposed function as vesicular adapter proteins in early stages of secretion. Neuron. 1997 Mar;18(3):453–461. [PubMed]
  • Wiser O, Bennett MK, Atlas D. Functional interaction of syntaxin and SNAP-25 with voltage-sensitive L- and N-type Ca2+ channels. EMBO J. 1996 Aug 15;15(16):4100–4110. [PMC free article] [PubMed]

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