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PMID: 8636207 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Biochemical requirements for the targeting and fusion of ER-derived transport vesicles with purified yeast Golgi membranes.

The Journal of cell biology ·Vol. 132 ·No. 3 ·1996-02-00 ·Pages 277-89

Lupashin VV, Hamamoto S, Schekman RW

Abstract

In order for secretion to progress, ER-derived transport vesicles must target to, and fuse with the cis-Golgi compartment. These processes have been reconstituted using highly enriched membrane fractions and partially purified soluble components. The functionally active yeast Golgi membranes that have been purified are highly enriched in the cis-Golgi marker enzymes alpha 1,6 mannosyltransferase and GDPase. Fusion of transport vesicles with these membranes requires both GTP and ATP hydrolysis, and depends on cytosolic and peripheral membrane proteins. At least two protein fractions from yeast cytosol are required for the reconstitution of ER-derived vesicle fusion. Soluble fractions prepared from temperature-sensitive mutants revealed requirements for the Ypt1p, Sec19p, Sly1p, Sec7p, and Uso1 proteins. A model for the sequential involvement of these components in the targeting and fusion reaction is proposed.

MeSH Terms
Biological Transport Biomarkers Cell Fractionation/methods Endoplasmic Reticulum/physiology,ultrastructure Fungal Proteins/analysis Golgi Apparatus/physiology,ultrastructure Intracellular Membranes/physiology,ultrastructure Mannosyltransferases/analysis Membrane Fusion Microscopy, Electron Organelles/physiology,ultrastructure Pyrophosphatases/analysis Saccharomyces cerevisiae/genetics,physiology,ultrastructure
Chemicals
Biomarkers Fungal Proteins Mannosyltransferases alpha 1,6-mannosyltransferase Pyrophosphatases guanosine-diphosphatase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lupashin V V
Department of Molecular and Cell Biology, University of California, Berkeley 94720, USA.
Hamamoto S
Schekman R W
References (44)
44 references, click to expand
  1. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  2. OCH1 encodes a novel membrane bound mannosyltransferase: outer chain elongation of asparagine-linked oligosaccharides.
    EMBO J. 1992 Jul;11(7):2511-9 PMID: 1628616
  3. alpha-D-Mannosidase of Saccharomyces cerevisiae. Characterization and modulation of activity.
    Biochim Biophys Acta. 1978 May 11;524(1):121-30 PMID: 350285
  4. The effects of vanadate on the plasma membrane ATPase of Neurospora crassa.
    J Biol Chem. 1979 Apr 25;254(8):2928-34 PMID: 155060
  5. Reconstitution of SEC gene product-dependent intercompartmental protein transport.
    Cell. 1988 Jul 29;54(3):335-44 PMID: 3293799
  6. SEC7 encodes an unusual, high molecular weight protein required for membrane traffic from the yeast Golgi apparatus.
    J Biol Chem. 1988 Aug 25;263(24):11711-7 PMID: 3042778
  7. Yeast prohormone processing enzyme (KEX2 gene product) is a Ca2+-dependent serine protease.
    Proc Natl Acad Sci U S A. 1989 Mar;86(5):1434-8 PMID: 2646633
  8. A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast.
    Nature. 1989 Jun 1;339(6223):355-9 PMID: 2657434
  9. The GTP-binding protein Ypt1 is required for transport in vitro: the Golgi apparatus is defective in ypt1 mutants.
    J Cell Biol. 1989 Sep;109(3):1015-22 PMID: 2504726
  10. 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):6935-9 PMID: 2476806
  11. Vesicle-mediated protein sorting.
    Annu Rev Biochem. 1992;61:471-516 PMID: 1497318
  12. A novel 115-kD peripheral membrane protein is required for intercisternal transport in the Golgi stack.
    J Cell Biol. 1992 Sep;118(5):1015-26 PMID: 1512287
  13. Characterization of the Saccharomyces Golgi complex through the cell cycle by immunoelectron microscopy.
    Mol Biol Cell. 1992 Jul;3(7):789-803 PMID: 1381247
  14. 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):513-21 PMID: 1400588
  15. SEC21 is a gene required for ER to Golgi protein transport that encodes a subunit of a yeast coatomer.
    Nature. 1992 Dec 10;360(6404):603-5 PMID: 1461285
  16. Reconstitution of transport from endoplasmic reticulum to Golgi complex using endoplasmic reticulum-enriched membrane fraction from yeast.
    Methods Enzymol. 1992;219:124-36 PMID: 1487986
  17. SNAP receptors implicated in vesicle targeting and fusion.
    Nature. 1993 Mar 25;362(6418):318-24 PMID: 8455717
  18. 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):735-45 PMID: 8500167
  19. 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):409-18 PMID: 8221884
  20. Friends and family: the role of the Rab GTPases in vesicular traffic.
    Cell. 1993 Nov 19;75(4):597-601 PMID: 8242735
  21. 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):2003-7 PMID: 8134339
  22. GDI1 encodes a GDP dissociation inhibitor that plays an essential role in the yeast secretory pathway.
    EMBO J. 1994 Apr 1;13(7):1718-28 PMID: 8157010
  23. Molecular characterization of the USO1 gene product which is essential for vesicular transport in Saccharomyces cerevisiae.
    Biochem Biophys Res Commun. 1994 Apr 15;200(1):647-53 PMID: 8166741
  24. COPII: a membrane coat formed by Sec proteins that drive vesicle budding from the endoplasmic reticulum.
    Cell. 1994 Jun 17;77(6):895-907 PMID: 8004676
  25. A rab protein is required for the assembly of SNARE complexes in the docking of transport vesicles.
    Cell. 1994 Sep 23;78(6):937-48 PMID: 7923363
  26. Purification and characterization of a late Golgi compartment from Saccharomyces cerevisiae.
    J Biol Chem. 1994 Nov 11;269(45):28106-17 PMID: 7961747
  27. Clathrin-dependent localization of alpha 1,3 mannosyltransferase to the Golgi complex of Saccharomyces cerevisiae.
    J Cell Biol. 1994 Nov;127(3):667-78 PMID: 7962051
  28. Mechanisms of intracellular protein transport.
    Nature. 1994 Nov 3;372(6501):55-63 PMID: 7969419
  29. 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):522-6 PMID: 7831323
  30. Transcytosis-associated protein (TAP)/p115 is a general fusion factor required for binding of vesicles to acceptor membranes.
    Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):527-31 PMID: 7831324
  31. Functional compartments of the yeast Golgi apparatus are defined by the sec7 mutation.
    EMBO J. 1989 Sep;8(9):2695-702 PMID: 2684655
  32. GTP-binding Ypt1 protein and Ca2+ function independently in a cell-free protein transport reaction.
    Proc Natl Acad Sci U S A. 1990 Jan;87(1):355-9 PMID: 2104983
  33. SNAPs, a family of NSF attachment proteins involved in intracellular membrane fusion in animals and yeast.
    Cell. 1990 May 18;61(4):709-21 PMID: 2111733
  34. Distinct sets of SEC genes govern transport vesicle formation and fusion early in the secretory pathway.
    Cell. 1990 May 18;61(4):723-33 PMID: 2188733
  35. 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):872-85 PMID: 1990290
  36. A cytoskeleton-related gene, uso1, is required for intracellular protein transport in Saccharomyces cerevisiae.
    J Cell Biol. 1991 Apr;113(2):245-60 PMID: 2010462
  37. Immunolocalization of Kex2 protease identifies a putative late Golgi compartment in the yeast Saccharomyces cerevisiae.
    J Cell Biol. 1991 May;113(3):527-38 PMID: 2016334
  38. The yeast SLY gene products, suppressors of defects in the essential GTP-binding Ypt1 protein, may act in endoplasmic reticulum-to-Golgi transport.
    Mol Cell Biol. 1991 Jun;11(6):2980-93 PMID: 1903839
  39. Mediation of the attachment or fusion step in vesicular transport by the GTP-binding Ypt1 protein.
    Science. 1991 Jun 14;252(5012):1553-6 PMID: 1904626
  40. Distinct biochemical requirements for the budding, targeting, and fusion of ER-derived transport vesicles.
    J Cell Biol. 1991 Jul;114(2):219-29 PMID: 1649197
  41. Immuno-isolation of Sec7p-coated transport vesicles from the yeast secretory pathway.
    Nature. 1992 Jan 9;355(6356):173-5 PMID: 1729652
  42. A multisubunit particle implicated in membrane fusion.
    J Cell Biol. 1992 May;117(3):531-8 PMID: 1315316
  43. The yeast SEC17 gene product is functionally equivalent to mammalian alpha-SNAP protein.
    J Biol Chem. 1992 Jun 15;267(17):12106-15 PMID: 1601878
  44. Cytochrome c oxidase from bakers' yeast. I. Isolation and properties.
    J Biol Chem. 1973 Feb 25;248(4):1346-54 PMID: 4346952
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.132.3.277
Published
1996-02-00
Pages
277-89
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120720
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.132.3.277 PMC Full Text View journal details
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