Skip to main content
NCBI home page
The EMBO Journal logoLink to The EMBO Journal
. 1997 Jun 2;16(11):3017–3024. doi: 10.1093/emboj/16.11.3017

A novel SNARE complex implicated in vesicle fusion with the endoplasmic reticulum.

M J Lewis 1, J C Rayner 1, H R Pelham 1
PMCID: PMC1169920  PMID: 9214619

Abstract

Intracellular vesicular traffic is controlled in part by v- and t-SNAREs, integral membrane proteins which allow specific interaction and fusion between vesicles (v-SNAREs) and their target membranes (t-SNAREs). In yeast, retrograde transport from the Golgi complex to the ER is mediated by the ER t-SNARE Ufe1p, and also requires two other ER proteins, Sec20p and Tip20p, which bind each other. Although Sec20p is not a typical SNARE, we show that both it and Tip20p can be co-precipitated with Ufe1p, and that a growth-inhibiting mutation in Ufe1p can be compensated by a mutation in Sec20p. Furthermore, Sec22p, a v-SNARE implicated in forward transport from ER to Golgi, co-precipitates with Ufe1p and Sec20p, and SEC22 acts as an allele-specific multicopy suppressor of a temperature-sensitive ufe1 mutation. These results define a new functional SNARE complex, with features distinct from the plasma membrane and cis-Golgi complexes previously identified. They also show that a single v-SNARE can be involved in both anterograde and retrograde transport, which suggests that the mere presence of a particular v-SNARE may not be sufficient to determine the preferred target for a transport vesicle.

Full Text

The Full Text of this article is available as a PDF (261.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Banfield D. K., Lewis M. J., Pelham H. R. A SNARE-like protein required for traffic through the Golgi complex. Nature. 1995 Jun 29;375(6534):806–809. doi: 10.1038/375806a0. [DOI] [PubMed] [Google Scholar]
  2. Barlowe C., Orci L., Yeung T., Hosobuchi M., Hamamoto S., Salama N., Rexach M. F., Ravazzola M., Amherdt M., Schekman R. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum. Cell. 1994 Jun 17;77(6):895–907. doi: 10.1016/0092-8674(94)90138-4. [DOI] [PubMed] [Google Scholar]
  3. Becherer K. A., Rieder S. E., Emr S. D., Jones E. W. Novel syntaxin homologue, Pep12p, required for the sorting of lumenal hydrolases to the lysosome-like vacuole in yeast. Mol Biol Cell. 1996 Apr;7(4):579–594. doi: 10.1091/mbc.7.4.579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eakle K. A., Bernstein M., Emr S. D. Characterization of a component of the yeast secretion machinery: identification of the SEC18 gene product. Mol Cell Biol. 1988 Oct;8(10):4098–4109. doi: 10.1128/mcb.8.10.4098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ferro-Novick S., Jahn R. Vesicle fusion from yeast to man. Nature. 1994 Jul 21;370(6486):191–193. doi: 10.1038/370191a0. [DOI] [PubMed] [Google Scholar]
  6. Fresko I., Yazici H., Bayramiçli M., Yurdakul S., Mat C. Effect of surgical cleaning of the skin on the pathergy phenomenon in Behçet's syndrome. Ann Rheum Dis. 1993 Aug;52(8):619–620. doi: 10.1136/ard.52.8.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Griff I. C., Schekman R., Rothman J. E., Kaiser C. A. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein. J Biol Chem. 1992 Jun 15;267(17):12106–12115. [PubMed] [Google Scholar]
  8. Hardwick K. G., Pelham H. R. SED5 encodes a 39-kD integral membrane protein required for vesicular transport between the ER and the Golgi complex. J Cell Biol. 1992 Nov;119(3):513–521. doi: 10.1083/jcb.119.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hayashi T., McMahon H., Yamasaki S., Binz T., Hata Y., Südhof T. C., Niemann H. Synaptic vesicle membrane fusion complex: action of clostridial neurotoxins on assembly. EMBO J. 1994 Nov 1;13(21):5051–5061. doi: 10.1002/j.1460-2075.1994.tb06834.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hayashi T., Yamasaki S., Nauenburg S., Binz T., Niemann H. Disassembly of the reconstituted synaptic vesicle membrane fusion complex in vitro. EMBO J. 1995 May 15;14(10):2317–2325. doi: 10.1002/j.1460-2075.1995.tb07226.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jiang Y., Sacher M., Singer-Krüger B., Lian J. P., Stone S., Ferro-Novick S. Factors mediating the late stages of ER-to-Golgi transport in yeast. Cold Spring Harb Symp Quant Biol. 1995;60:119–126. doi: 10.1101/sqb.1995.060.01.015. [DOI] [PubMed] [Google Scholar]
  12. Kaiser C. A., Schekman R. Distinct sets of SEC genes govern transport vesicle formation and fusion early in the secretory pathway. Cell. 1990 May 18;61(4):723–733. doi: 10.1016/0092-8674(90)90483-u. [DOI] [PubMed] [Google Scholar]
  13. Kee Y., Lin R. C., Hsu S. C., Scheller R. H. Distinct domains of syntaxin are required for synaptic vesicle fusion complex formation and dissociation. Neuron. 1995 May;14(5):991–998. doi: 10.1016/0896-6273(95)90337-2. [DOI] [PubMed] [Google Scholar]
  14. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  15. Letourneur F., Gaynor E. C., Hennecke S., Démollière C., Duden R., Emr S. D., Riezman H., Cosson P. Coatomer is essential for retrieval of dilysine-tagged proteins to the endoplasmic reticulum. Cell. 1994 Dec 30;79(7):1199–1207. doi: 10.1016/0092-8674(94)90011-6. [DOI] [PubMed] [Google Scholar]
  16. Lewis M. J., Pelham H. R. SNARE-mediated retrograde traffic from the Golgi complex to the endoplasmic reticulum. Cell. 1996 Apr 19;85(2):205–215. doi: 10.1016/s0092-8674(00)81097-1. [DOI] [PubMed] [Google Scholar]
  17. Lian J. P., Ferro-Novick S. Bos1p, an integral membrane protein of the endoplasmic reticulum to Golgi transport vesicles, is required for their fusion competence. Cell. 1993 May 21;73(4):735–745. doi: 10.1016/0092-8674(93)90253-m. [DOI] [PubMed] [Google Scholar]
  18. Lian J. P., Stone S., Jiang Y., Lyons P., Ferro-Novick S. Ypt1p implicated in v-SNARE activation. Nature. 1994 Dec 15;372(6507):698–701. doi: 10.1038/372698a0. [DOI] [PubMed] [Google Scholar]
  19. McMahon H. T., Südhof T. C. Synaptic core complex of synaptobrevin, syntaxin, and SNAP25 forms high affinity alpha-SNAP binding site. J Biol Chem. 1995 Feb 3;270(5):2213–2217. doi: 10.1074/jbc.270.5.2213. [DOI] [PubMed] [Google Scholar]
  20. Munro S., Pelham H. R. A C-terminal signal prevents secretion of luminal ER proteins. Cell. 1987 Mar 13;48(5):899–907. doi: 10.1016/0092-8674(87)90086-9. [DOI] [PubMed] [Google Scholar]
  21. Nagahama M., Orci L., Ravazzola M., Amherdt M., Lacomis L., Tempst P., Rothman J. E., Söllner T. H. A v-SNARE implicated in intra-Golgi transport. J Cell Biol. 1996 May;133(3):507–516. doi: 10.1083/jcb.133.3.507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pevsner J., Hsu S. C., Braun J. E., Calakos N., Ting A. E., Bennett M. K., Scheller R. H. Specificity and regulation of a synaptic vesicle docking complex. Neuron. 1994 Aug;13(2):353–361. doi: 10.1016/0896-6273(94)90352-2. [DOI] [PubMed] [Google Scholar]
  23. Pevsner J., Hsu S. C., Braun J. E., Calakos N., Ting A. E., Bennett M. K., Scheller R. H. Specificity and regulation of a synaptic vesicle docking complex. Neuron. 1994 Aug;13(2):353–361. doi: 10.1016/0896-6273(94)90352-2. [DOI] [PubMed] [Google Scholar]
  24. Protopopov V., Govindan B., Novick P., Gerst J. E. 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. doi: 10.1016/0092-8674(93)90465-3. [DOI] [PubMed] [Google Scholar]
  25. Rose M. D., Misra L. M., Vogel J. P. KAR2, a karyogamy gene, is the yeast homolog of the mammalian BiP/GRP78 gene. Cell. 1989 Jun 30;57(7):1211–1221. doi: 10.1016/0092-8674(89)90058-5. [DOI] [PubMed] [Google Scholar]
  26. Semenza J. C., Hardwick K. G., Dean N., Pelham H. R. ERD2, a yeast gene required for the receptor-mediated retrieval of luminal ER proteins from the secretory pathway. Cell. 1990 Jun 29;61(7):1349–1357. doi: 10.1016/0092-8674(90)90698-e. [DOI] [PubMed] [Google Scholar]
  27. Sikorski R. S., Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. doi: 10.1093/genetics/122.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Subramaniam V. N., Peter F., Philp R., Wong S. H., Hong W. GS28, a 28-kilodalton Golgi SNARE that participates in ER-Golgi transport. Science. 1996 May 24;272(5265):1161–1163. doi: 10.1126/science.272.5265.1161. [DOI] [PubMed] [Google Scholar]
  29. Sweet D. J., Pelham H. R. The Saccharomyces cerevisiae SEC20 gene encodes a membrane glycoprotein which is sorted by the HDEL retrieval system. EMBO J. 1992 Feb;11(2):423–432. doi: 10.1002/j.1460-2075.1992.tb05071.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sweet D. J., Pelham H. R. The TIP1 gene of Saccharomyces cerevisiae encodes an 80 kDa cytoplasmic protein that interacts with the cytoplasmic domain of Sec20p. EMBO J. 1993 Jul;12(7):2831–2840. doi: 10.1002/j.1460-2075.1993.tb05944.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Söllner T., Bennett M. K., Whiteheart S. W., Scheller R. H., Rothman J. E. A protein assembly-disassembly pathway in vitro that may correspond to sequential steps of synaptic vesicle docking, activation, and fusion. Cell. 1993 Nov 5;75(3):409–418. doi: 10.1016/0092-8674(93)90376-2. [DOI] [PubMed] [Google Scholar]
  32. Söllner T., Whiteheart S. W., Brunner M., Erdjument-Bromage H., Geromanos S., Tempst P., Rothman J. E. SNAP receptors implicated in vesicle targeting and fusion. Nature. 1993 Mar 25;362(6418):318–324. doi: 10.1038/362318a0. [DOI] [PubMed] [Google Scholar]
  33. Søgaard M., Tani K., Ye R. R., Geromanos S., Tempst P., Kirchhausen T., Rothman J. E., Söllner T. A rab protein is required for the assembly of SNARE complexes in the docking of transport vesicles. Cell. 1994 Sep 23;78(6):937–948. doi: 10.1016/0092-8674(94)90270-4. [DOI] [PubMed] [Google Scholar]
  34. Way M., Pope B., Gooch J., Hawkins M., Weeds A. G. Identification of a region in segment 1 of gelsolin critical for actin binding. EMBO J. 1990 Dec;9(12):4103–4109. doi: 10.1002/j.1460-2075.1990.tb07632.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wilson D. W., Wilcox C. A., Flynn G. C., Chen E., Kuang W. J., Henzel W. J., Block M. R., Ullrich A., Rothman J. E. A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast. Nature. 1989 Jun 1;339(6223):355–359. doi: 10.1038/339355a0. [DOI] [PubMed] [Google Scholar]
  36. Wuestehube L. J., Duden R., Eun A., Hamamoto S., Korn P., Ram R., Schekman R. New mutants of Saccharomyces cerevisiae affected in the transport of proteins from the endoplasmic reticulum to the Golgi complex. Genetics. 1996 Feb;142(2):393–406. doi: 10.1093/genetics/142.2.393. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES