pmc logo image
Logo of embojJournal URL: http://www.nature.com/emboj/index.html
Journal List > EMBO J > v.17(9); May 1, 1998

Formats:
EMBO J. 1998 May 1; 17(9): 2494–2503.
doi: 10.1093/emboj/17.9.2494.
PMCID: PMC1170591
Copyright notice
TRAPP, a highly conserved novel complex on the cis-Golgi that mediates vesicle docking and fusion.
M Sacher, Y Jiang, J Barrowman, A Scarpa, J Burston, L Zhang, D Schieltz, J R Yates, 3rd, H Abeliovich, and S Ferro-Novick
Howard Hughes Medical Institute and the Department of Cell Biology, Yale University School of Medicine, New Haven, CT 06510, USA.
Small right arrow pointing to: This article has been cited by other articles in PMC.
Abstract
We previously identified BET3 by its genetic interactions with BET1, a gene whose SNARE-like product acts in endoplasmic reticulum (ER)-to-Golgi transport. To gain insight into the function of Bet3p, we added three c-myc tags to its C-terminus and immunopurified this protein from a clarified detergent extract. Here we report that Bet3p is a member of a large complex ( approximately 800 kDa) that we call TRAPP (transport protein particle). We propose that TRAPP plays a key role in the targeting and/or fusion of ER-to-Golgi transport vesicles with their acceptor compartment. The localization of Bet3p to the cis-Golgi complex, as well as biochemical studies showing that Bet3p functions on this compartment, support this hypothesis. TRAPP contains at least nine other constituents, five of which have been identified and shown to be highly conserved novel proteins.
Full Text
The Full Text of this article is available as a PDF (655K).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
  • Abeijon C, Orlean P, Robbins PW, Hirschberg CB. Topography of glycosylation in yeast: characterization of GDPmannose transport and lumenal guanosine diphosphatase activities in Golgi-like vesicles. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6935–6939. [PubMed]
  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • Banfield DK, Lewis MJ, Rabouille C, Warren G, Pelham HR. Localization of Sed5, a putative vesicle targeting molecule, to the cis-Golgi network involves both its transmembrane and cytoplasmic domains. J Cell Biol. 1994 Oct;127(2):357–371. [PubMed]
  • Barlowe C. Coupled ER to Golgi transport reconstituted with purified cytosolic proteins. J Cell Biol. 1997 Dec 1;139(5):1097–1108. [PubMed]
  • Brennwald P, Novick P. Interactions of three domains distinguishing the Ras-related GTP-binding proteins Ypt1 and Sec4. Nature. 1993 Apr 8;362(6420):560–563. [PubMed]
  • Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. [PubMed]
  • Cunningham KW, Wickner WT. Yeast KEX2 protease and mannosyltransferase I are localized to distinct compartments of the secretory pathway. Yeast. 1989 Jan–Feb;5(1):25–33. [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. [PubMed]
  • Ferro-Novick S, Jahn R. Vesicle fusion from yeast to man. Nature. 1994 Jul 21;370(6486):191–193. [PubMed]
  • von Mollard GF, Nothwehr SF, Stevens TH. The yeast v-SNARE Vti1p mediates two vesicle transport pathways through interactions with the t-SNAREs Sed5p and Pep12p. J Cell Biol. 1997 Jun 30;137(7):1511–1524. [PubMed]
  • Griff IC, Schekman R, Rothman JE, Kaiser CA. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein. J Biol Chem. 1992 Jun 15;267(17):12106–12115. [PubMed]
  • Groesch ME, Ruohola H, Bacon R, Rossi G, Ferro-Novick S. Isolation of a functional vesicular intermediate that mediates ER to Golgi transport in yeast. J Cell Biol. 1990 Jul;111(1):45–53. [PubMed]
  • Guo W, Roth D, Gatti E, De Camilli P, Novick P. Identification and characterization of homologues of the Exocyst component Sec10p. FEBS Lett. 1997 Mar 10;404(2-3):135–139. [PubMed]
  • Hardwick KG, Pelham HR. 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. [PubMed]
  • Hsu SC, Ting AE, Hazuka CD, Davanger S, Kenny JW, Kee Y, Scheller RH. The mammalian brain rsec6/8 complex. Neuron. 1996 Dec;17(6):1209–1219. [PubMed]
  • Lian JP, 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. [PubMed]
  • Lian JP, Stone S, Jiang Y, Lyons P, Ferro-Novick S. Ypt1p implicated in v-SNARE activation. Nature. 1994 Dec 15;372(6507):698–701. [PubMed]
  • Lupas A, Van Dyke M, Stock J. Predicting coiled coils from protein sequences. Science. 1991 May 24;252(5010):1162–1164. [PubMed]
  • Mayer A, Wickner W, Haas A. Sec18p (NSF)-driven release of Sec17p (alpha-SNAP) can precede docking and fusion of yeast vacuoles. Cell. 1996 Apr 5;85(1):83–94. [PubMed]
  • McCormack AL, Schieltz DM, Goode B, Yang S, Barnes G, Drubin D, Yates JR., 3rd Direct analysis and identification of proteins in mixtures by LC/MS/MS and database searching at the low-femtomole level. Anal Chem. 1997 Feb 15;69(4):767–776. [PubMed]
  • Nakajima H, Hirata A, Ogawa Y, Yonehara T, Yoda K, Yamasaki M. A cytoskeleton-related gene, uso1, is required for intracellular protein transport in Saccharomyces cerevisiae. J Cell Biol. 1991 Apr;113(2):245–260. [PubMed]
  • Newman AP, Groesch ME, Ferro-Novick S. Bos1p, a membrane protein required for ER to Golgi transport in yeast, co-purifies with the carrier vesicles and with Bet1p and the ER membrane. EMBO J. 1992 Oct;11(10):3609–3617. [PubMed]
  • Novick P, Ferro S, Schekman R. Order of events in the yeast secretory pathway. Cell. 1981 Aug;25(2):461–469. [PubMed]
  • Prasher DC, Eckenrode VK, Ward WW, Prendergast FG, Cormier MJ. Primary structure of the Aequorea victoria green-fluorescent protein. Gene. 1992 Feb 15;111(2):229–233. [PubMed]
  • Rose MD, Misra LM, Vogel JP. KAR2, a karyogamy gene, is the yeast homolog of the mammalian BiP/GRP78 gene. Cell. 1989 Jun 30;57(7):1211–1221. [PubMed]
  • Rossi G, Kolstad K, Stone S, Palluault F, Ferro-Novick S. BET3 encodes a novel hydrophilic protein that acts in conjunction with yeast SNAREs. Mol Biol Cell. 1995 Dec;6(12):1769–1780. [PubMed]
  • Ruohola H, Kabcenell AK, Ferro-Novick S. Reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex in yeast: the acceptor Golgi compartment is defective in the sec23 mutant. J Cell Biol. 1988 Oct;107(4):1465–1476. [PubMed]
  • Sacher M, Stone S, Ferro-Novick S. The synaptobrevin-related domains of Bos1p and Sec22p bind to the syntaxin-like region of Sed5p. J Biol Chem. 1997 Jul 4;272(27):17134–17138. [PubMed]
  • Sapperstein SK, Lupashin VV, Schmitt HD, Waters MG. Assembly of the ER to Golgi SNARE complex requires Uso1p. J Cell Biol. 1996 Mar;132(5):755–767. [PubMed]
  • Segev N, Mulholland J, Botstein D. The yeast GTP-binding YPT1 protein and a mammalian counterpart are associated with the secretion machinery. Cell. 1988 Mar 25;52(6):915–924. [PubMed]
  • Singer-Krüger B, Ferro-Novick S. Use of a synthetic lethal screen to identify yeast mutants impaired in endocytosis, vacuolar protein sorting and the organization of the cytoskeleton. Eur J Cell Biol. 1997 Dec;74(4):365–375. [PubMed]
  • Søgaard M, Tani K, Ye RR, Geromanos S, Tempst P, Kirchhausen T, Rothman JE, 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. [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]
  • Söllner T, Bennett MK, Whiteheart SW, Scheller RH, Rothman JE. 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. [PubMed]
  • Stone S, Sacher M, Mao Y, Carr C, Lyons P, Quinn AM, Ferro-Novick S. Bet1p activates the v-SNARE Bos1p. Mol Biol Cell. 1997 Jul;8(7):1175–1181. [PubMed]
  • TerBush DR, Novick P. Sec6, Sec8, and Sec15 are components of a multisubunit complex which localizes to small bud tips in Saccharomyces cerevisiae. J Cell Biol. 1995 Jul;130(2):299–312. [PubMed]
  • TerBush DR, Maurice T, Roth D, Novick P. The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae. EMBO J. 1996 Dec 2;15(23):6483–6494. [PubMed]
  • Wilson DW, Wilcox CA, Flynn GC, Chen E, Kuang WJ, Henzel WJ, Block MR, Ullrich A, Rothman JE. A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast. Nature. 1989 Jun 1;339(6223):355–359. [PubMed]
  • Yates JR, 3rd, Eng JK, McCormack AL, Schieltz D. Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database. Anal Chem. 1995 Apr 15;67(8):1426–1436. [PubMed]
Articles from The EMBO Journal are provided here courtesy of
The European Molecular Biology Organization