Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1999 Feb 15;18(4):1071–1080. doi: 10.1093/emboj/18.4.1071

The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis.

W Guo 1, D Roth 1, C Walch-Solimena 1, P Novick 1
PMCID: PMC1171198  PMID: 10022848

Abstract

Polarized secretion requires proper targeting of secretory vesicles to specific sites on the plasma membrane. Here we report that the exocyst complex plays a key role in vesicle targeting. Sec15p, an exocyst component, can associate with secretory vesicles and interact specifically with the rab GTPase, Sec4p, in its GTP-bound form. A chain of protein-protein interactions leads from Sec4p and Sec15p on the vesicle, through various subunits of the exocyst, to Sec3p, which marks the sites of exocytosis on the plasma membrane. Sec4p may control the assembly of the exocyst. The exocyst may therefore function as a rab effector system for targeted secretion.

Full Text

The Full Text of this article is available as a PDF (457.3 KB).

Selected References

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

  1. Ayscough K. R., Stryker J., Pokala N., Sanders M., Crews P., Drubin D. G. High rates of actin filament turnover in budding yeast and roles for actin in establishment and maintenance of cell polarity revealed using the actin inhibitor latrunculin-A. J Cell Biol. 1997 Apr 21;137(2):399–416. doi: 10.1083/jcb.137.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brennwald P., Kearns B., Champion K., Keränen S., Bankaitis V., Novick P. Sec9 is a SNAP-25-like component of a yeast SNARE complex that may be the effector of Sec4 function in exocytosis. Cell. 1994 Oct 21;79(2):245–258. doi: 10.1016/0092-8674(94)90194-5. [DOI] [PubMed] [Google Scholar]
  3. 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. doi: 10.1038/362560a0. [DOI] [PubMed] [Google Scholar]
  4. Cao X., Ballew N., Barlowe C. Initial docking of ER-derived vesicles requires Uso1p and Ypt1p but is independent of SNARE proteins. EMBO J. 1998 Apr 15;17(8):2156–2165. doi: 10.1093/emboj/17.8.2156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chien C. T., Bartel P. L., Sternglanz R., Fields S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9578–9582. doi: 10.1073/pnas.88.21.9578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Collins R. N., Brennwald P., Garrett M., Lauring A., Novick P. Interactions of nucleotide release factor Dss4p with Sec4p in the post-Golgi secretory pathway of yeast. J Biol Chem. 1997 Jul 18;272(29):18281–18289. doi: 10.1074/jbc.272.29.18281. [DOI] [PubMed] [Google Scholar]
  7. Cunningham K. W., Wickner W. T. Yeast KEX2 protease and mannosyltransferase I are localized to distinct compartments of the secretory pathway. Yeast. 1989 Jan-Feb;5(1):25–33. doi: 10.1002/yea.320050105. [DOI] [PubMed] [Google Scholar]
  8. Durfee T., Becherer K., Chen P. L., Yeh S. H., Yang Y., Kilburn A. E., Lee W. H., Elledge S. J. The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. Genes Dev. 1993 Apr;7(4):555–569. doi: 10.1101/gad.7.4.555. [DOI] [PubMed] [Google Scholar]
  9. Díaz E., Schimmöller F., Pfeffer S. R. A novel Rab9 effector required for endosome-to-TGN transport. J Cell Biol. 1997 Jul 28;138(2):283–290. doi: 10.1083/jcb.138.2.283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Echard A., Jollivet F., Martinez O., Lacapère J. J., Rousselet A., Janoueix-Lerosey I., Goud B. Interaction of a Golgi-associated kinesin-like protein with Rab6. Science. 1998 Jan 23;279(5350):580–585. doi: 10.1126/science.279.5350.580. [DOI] [PubMed] [Google Scholar]
  11. Finger F. P., Hughes T. E., Novick P. Sec3p is a spatial landmark for polarized secretion in budding yeast. Cell. 1998 Feb 20;92(4):559–571. doi: 10.1016/s0092-8674(00)80948-4. [DOI] [PubMed] [Google Scholar]
  12. Garrett M. D., Zahner J. E., Cheney C. M., Novick P. J. GDI1 encodes a GDP dissociation inhibitor that plays an essential role in the yeast secretory pathway. EMBO J. 1994 Apr 1;13(7):1718–1728. doi: 10.1002/j.1460-2075.1994.tb06436.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goud B., Salminen A., Walworth N. C., Novick P. J. A GTP-binding protein required for secretion rapidly associates with secretory vesicles and the plasma membrane in yeast. Cell. 1988 Jun 3;53(5):753–768. doi: 10.1016/0092-8674(88)90093-1. [DOI] [PubMed] [Google Scholar]
  14. Govindan B., Bowser R., Novick P. The role of Myo2, a yeast class V myosin, in vesicular transport. J Cell Biol. 1995 Mar;128(6):1055–1068. doi: 10.1083/jcb.128.6.1055. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grindstaff K. K., Yeaman C., Anandasabapathy N., Hsu S. C., Rodriguez-Boulan E., Scheller R. H., Nelson W. J. Sec6/8 complex is recruited to cell-cell contacts and specifies transport vesicle delivery to the basal-lateral membrane in epithelial cells. Cell. 1998 May 29;93(5):731–740. doi: 10.1016/s0092-8674(00)81435-x. [DOI] [PubMed] [Google Scholar]
  16. 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. doi: 10.1016/s0014-5793(97)00109-9. [DOI] [PubMed] [Google Scholar]
  17. Hazuka C. D., Hsu S. C., Scheller R. H. Characterization of a cDNA encoding a subunit of the rat brain rsec6/8 complex. Gene. 1997 Mar 10;187(1):67–73. doi: 10.1016/s0378-1119(96)00720-2. [DOI] [PubMed] [Google Scholar]
  18. Hsu S. C., Ting A. E., Hazuka C. D., Davanger S., Kenny J. W., Kee Y., Scheller R. H. The mammalian brain rsec6/8 complex. Neuron. 1996 Dec;17(6):1209–1219. doi: 10.1016/s0896-6273(00)80251-2. [DOI] [PubMed] [Google Scholar]
  19. Jiang Y., Rossi G., Ferro-Novick S. Bet2p and Mad2p are components of a prenyltransferase that adds geranylgeranyl onto Ypt1p and Sec4p. Nature. 1993 Nov 4;366(6450):84–86. doi: 10.1038/366084a0. [DOI] [PubMed] [Google Scholar]
  20. Kee Y., Yoo J. S., Hazuka C. D., Peterson K. E., Hsu S. C., Scheller R. H. Subunit structure of the mammalian exocyst complex. Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14438–14443. doi: 10.1073/pnas.94.26.14438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Mondésert G., Clarke D. J., Reed S. I. Identification of genes controlling growth polarity in the budding yeast Saccharomyces cerevisiae: a possible role of N-glycosylation and involvement of the exocyst complex. Genetics. 1997 Oct;147(2):421–434. doi: 10.1093/genetics/147.2.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mulholland J., Preuss D., Moon A., Wong A., Drubin D., Botstein D. Ultrastructure of the yeast actin cytoskeleton and its association with the plasma membrane. J Cell Biol. 1994 Apr;125(2):381–391. doi: 10.1083/jcb.125.2.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nakamura N., Lowe M., Levine T. P., Rabouille C., Warren G. The vesicle docking protein p115 binds GM130, a cis-Golgi matrix protein, in a mitotically regulated manner. Cell. 1997 May 2;89(3):445–455. doi: 10.1016/s0092-8674(00)80225-1. [DOI] [PubMed] [Google Scholar]
  24. Pfeffer S. R. Transport vesicle docking: SNAREs and associates. Annu Rev Cell Dev Biol. 1996;12:441–461. doi: 10.1146/annurev.cellbio.12.1.441. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Redding K., Holcomb C., Fuller R. S. Immunolocalization of Kex2 protease identifies a putative late Golgi compartment in the yeast Saccharomyces cerevisiae. J Cell Biol. 1991 May;113(3):527–538. doi: 10.1083/jcb.113.3.527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ren M., Zeng J., De Lemos-Chiarandini C., Rosenfeld M., Adesnik M., Sabatini D. D. In its active form, the GTP-binding protein rab8 interacts with a stress-activated protein kinase. Proc Natl Acad Sci U S A. 1996 May 14;93(10):5151–5155. doi: 10.1073/pnas.93.10.5151. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rieder S. E., Emr S. D. A novel RING finger protein complex essential for a late step in protein transport to the yeast vacuole. Mol Biol Cell. 1997 Nov;8(11):2307–2327. doi: 10.1091/mbc.8.11.2307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Roth D., Guo W., Novick P. Dominant negative alleles of SEC10 reveal distinct domains involved in secretion and morphogenesis in yeast. Mol Biol Cell. 1998 Jul;9(7):1725–1739. doi: 10.1091/mbc.9.7.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sacher M., Jiang Y., Barrowman J., Scarpa A., Burston J., Zhang L., Schieltz D., Yates J. R., 3rd, Abeliovich H., Ferro-Novick S. TRAPP, a highly conserved novel complex on the cis-Golgi that mediates vesicle docking and fusion. EMBO J. 1998 May 1;17(9):2494–2503. doi: 10.1093/emboj/17.9.2494. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Salminen A., Novick P. J. The Sec15 protein responds to the function of the GTP binding protein, Sec4, to control vesicular traffic in yeast. J Cell Biol. 1989 Sep;109(3):1023–1036. doi: 10.1083/jcb.109.3.1023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sapperstein S. K., Lupashin V. V., Schmitt H. D., Waters M. G. Assembly of the ER to Golgi SNARE complex requires Uso1p. J Cell Biol. 1996 Mar;132(5):755–767. doi: 10.1083/jcb.132.5.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sapperstein S. K., Walter D. M., Grosvenor A. R., Heuser J. E., Waters M. G. p115 is a general vesicular transport factor related to the yeast endoplasmic reticulum to Golgi transport factor Uso1p. Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):522–526. doi: 10.1073/pnas.92.2.522. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Sasaki T., Kikuchi A., Araki S., Hata Y., Isomura M., Kuroda S., Takai Y. Purification and characterization from bovine brain cytosol of a protein that inhibits the dissociation of GDP from and the subsequent binding of GTP to smg p25A, a ras p21-like GTP-binding protein. J Biol Chem. 1990 Feb 5;265(4):2333–2337. [PubMed] [Google Scholar]
  35. Schneider B. L., Seufert W., Steiner B., Yang Q. H., Futcher A. B. Use of polymerase chain reaction epitope tagging for protein tagging in Saccharomyces cerevisiae. Yeast. 1995 Oct;11(13):1265–1274. doi: 10.1002/yea.320111306. [DOI] [PubMed] [Google Scholar]
  36. Shirataki H., Kaibuchi K., Sakoda T., Kishida S., Yamaguchi T., Wada K., Miyazaki M., Takai Y. Rabphilin-3A, a putative target protein for smg p25A/rab3A p25 small GTP-binding protein related to synaptotagmin. Mol Cell Biol. 1993 Apr;13(4):2061–2068. doi: 10.1128/mcb.13.4.2061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Simonsen A., Lippé R., Christoforidis S., Gaullier J. M., Brech A., Callaghan J., Toh B. H., Murphy C., Zerial M., Stenmark H. EEA1 links PI(3)K function to Rab5 regulation of endosome fusion. Nature. 1998 Jul 30;394(6692):494–498. doi: 10.1038/28879. [DOI] [PubMed] [Google Scholar]
  38. Stenmark H., Vitale G., Ullrich O., Zerial M. Rabaptin-5 is a direct effector of the small GTPase Rab5 in endocytic membrane fusion. Cell. 1995 Nov 3;83(3):423–432. doi: 10.1016/0092-8674(95)90120-5. [DOI] [PubMed] [Google Scholar]
  39. Sönnichsen B., Lowe M., Levine T., Jämsä E., Dirac-Svejstrup B., Warren G. A role for giantin in docking COPI vesicles to Golgi membranes. J Cell Biol. 1998 Mar 9;140(5):1013–1021. doi: 10.1083/jcb.140.5.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. TerBush D. R., 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. [PMC free article] [PubMed] [Google Scholar]
  41. TerBush D. R., 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. doi: 10.1083/jcb.130.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Ting A. E., Hazuka C. D., Hsu S. C., Kirk M. D., Bean A. J., Scheller R. H. rSec6 and rSec8, mammalian homologs of yeast proteins essential for secretion. Proc Natl Acad Sci U S A. 1995 Oct 10;92(21):9613–9617. doi: 10.1073/pnas.92.21.9613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Verhage M., de Vries K. J., Røshol H., Burbach J. P., Gispen W. H., Südhof T. C. 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. doi: 10.1016/s0896-6273(00)81245-3. [DOI] [PubMed] [Google Scholar]
  44. Walch-Solimena C., Collins R. N., Novick P. J. Sec2p mediates nucleotide exchange on Sec4p and is involved in polarized delivery of post-Golgi vesicles. J Cell Biol. 1997 Jun 30;137(7):1495–1509. doi: 10.1083/jcb.137.7.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Walworth N. C., Brennwald P., Kabcenell A. K., Garrett M., Novick P. Hydrolysis of GTP by Sec4 protein plays an important role in vesicular transport and is stimulated by a GTPase-activating protein in Saccharomyces cerevisiae. Mol Cell Biol. 1992 May;12(5):2017–2028. doi: 10.1128/mcb.12.5.2017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Walworth N. C., Goud B., Kabcenell A. K., Novick P. J. Mutational analysis of SEC4 suggests a cyclical mechanism for the regulation of vesicular traffic. EMBO J. 1989 Jun;8(6):1685–1693. doi: 10.1002/j.1460-2075.1989.tb03560.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wang Y., Okamoto M., Schmitz F., Hofmann K., Südhof T. C. Rim is a putative Rab3 effector in regulating synaptic-vesicle fusion. Nature. 1997 Aug 7;388(6642):593–598. doi: 10.1038/41580. [DOI] [PubMed] [Google Scholar]

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

RESOURCES