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Vol. 11, Issue 1, 23-38, January 2000

Specific Retrieval of the Exocytic SNARE Snc1p from Early Yeast Endosomes

Michael J. Lewis,* Benjamin J. Nichols,*dagger Cristina Prescianotto-Baschong,Dagger Howard Riezman,Dagger and Hugh R. B. Pelham*§

 *Medical Research Council Laboratory of Molecular Biology, Cambridge CB2 2QH, United Kingdom; and  Dagger Biozentrum of the University of Basel, CH-4056 Basel, Switzerland

Many endocytosed proteins in yeast travel to the vacuole, but some are recycled to the plasma membrane. We have investigated the recycling of chimeras containing green fluorescent protein (GFP) and the exocytic SNARE Snc1p. GFP-Snc1p moves from the cell surface to internal structures when Golgi function or exocytosis is blocked, suggesting continuous recycling via the Golgi. Internalization is mediated by a conserved cytoplasmic signal, whereas diversion from the vacuolar pathway requires sequences within and adjacent to the transmembrane domain. Delivery from the Golgi to the surface is also influenced by the transmembrane domain, but the requirements are much less specific. Recycling requires the syntaxins Tlg1p and Tlg2p but not Pep12p or proteins such as Vps4p and Vps5p that have been implicated in late endosome-Golgi traffic. Subtle changes to the recycling signal cause GFP-Snc1p to accumulate preferentially in punctate internal structures, although it continues to recycle to the surface. The internal GFP-Snc1p colocalizes with Tlg1p, and immunofluorescence and immunoelectron microscopy reveal structures that contain Tlg1p, Tlg2p, and Kex2p but lack Pep12p and Sec7p. We propose that these represent early endosomes in which sorting of Snc1p and late Golgi proteins occurs, and that transport can occur directly from them to the Golgi apparatus.


dagger Present address: Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.

§ Corresponding author. E-mail address: hp{at}mrc-lmb.cam.ac.uk.


Molecular Biology of the Cell
Vol. 11, 23-38, January 2000
Copyright © 2000 by The American Society for Cell Biology



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J. Cell Biol., December 10, 2001; 155(6): 969 - 978.
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Mol. Biol. CellHome page
A. A. Sanderfoot, V. Kovaleva, D. C. Bassham, and N. V. Raikhel
Interactions between Syntaxins Identify at Least Five SNARE Complexes within the Golgi/Prevacuolar System of the Arabidopsis Cell
Mol. Biol. Cell, December 1, 2001; 12(12): 3733 - 3743.
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J. Biol. Chem.Home page
J.-O. De Craene, O. Soetens, and B. Andre
The Npr1 Kinase Controls Biosynthetic and Endocytic Sorting of the Yeast Gap1 Permease
J. Biol. Chem., November 16, 2001; 276(47): 43939 - 43948.
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Mol. Biol. CellHome page
S.-A. Ha, J. T. Bunch, H. Hama, D. B. DeWald, and S. F. Nothwehr
A Novel Mechanism for Localizing Membrane Proteins to Yeast Trans-Golgi Network Requires Function of Synaptojanin-like Protein
Mol. Biol. Cell, October 1, 2001; 12(10): 3175 - 3190.
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Mol. Cell. Biol.Home page
S. Dupre and R. Haguenauer-Tsapis
Deubiquitination Step in the Endocytic Pathway of Yeast Plasma Membrane Proteins: Crucial Role of Doa4p Ubiquitin Isopeptidase
Mol. Cell. Biol., July 15, 2001; 21(14): 4482 - 4494.
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Mol. Biol. CellHome page
T. P. Levine and S. Munro
Dual Targeting of Osh1p, a Yeast Homologue of Oxysterol-binding Protein, to both the Golgi and the Nucleus-Vacuole Junction
Mol. Biol. Cell, June 1, 2001; 12(6): 1633 - 1644.
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Mol. Cell. Biol.Home page
J.-M. Galan, A. Wiederkehr, J. H. Seol, R. Haguenauer-Tsapis, R. J. Deshaies, H. Riezman, and M. Peter
Skp1p and the F-Box Protein Rcy1p Form a Non-SCF Complex Involved in Recycling of the SNARE Snc1p in Yeast
Mol. Cell. Biol., May 1, 2001; 21(9): 3105 - 3117.
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JCBHome page
O. W. Rossanese, C. A. Reinke, B. J. Bevis, A. T. Hammond, I. B. Sears, J. O'Connor, and B. S. Glick
A Role for Actin, Cdc1p, and Myo2p in the Inheritance of Late Golgi Elements in Saccharomyces cerevisiae
J. Cell Biol., March 26, 2001; 153(1): 47 - 62.
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Mol. Biol. CellHome page
N. Yahara, T. Ueda, K. Sato, and A. Nakano
Multiple Roles of Arf1 GTPase in the Yeast Exocytic and Endocytic Pathways
Mol. Biol. Cell, January 1, 2001; 12(1): 221 - 238.
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Mol. Biol. CellHome page
E. Grote, G. Vlacich, M. Pypaert, and P. J. Novick
A snc1 Endocytosis Mutant: Phenotypic Analysis and Suppression by Overproduction of Dihydrosphingosine Phosphate Lyase
Mol. Biol. Cell, December 1, 2000; 11(12): 4051 - 4065.
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Mol. Biol. CellHome page
F. Reggiori, M. W. Black, and H. R. B. Pelham
Polar Transmembrane Domains Target Proteins to the Interior of the Yeast Vacuole
Mol. Biol. Cell, November 1, 2000; 11(11): 3737 - 3749.
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M. Black and H. Pelham
A Selective Transport Route from Golgi to Late Endosomes that Requires the Yeast GGA Proteins
J. Cell Biol., October 30, 2000; 151(3): 587 - 600.
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Chen and N. Davis
Recycling of the Yeast a-Factor Receptor
J. Cell Biol., October 30, 2000; 151(3): 731 - 738.
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E. Grote, C. M. Carr, and P. J. Novick
Ordering the Final Events in Yeast Exocytosis
J. Cell Biol., October 18, 2000; 151(2): 439 - 452.
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Mol. Biol. CellHome page
S. Gurunathan, D. Chapman-Shimshoni, S. Trajkovic, and J. E. Gerst
Yeast Exocytic v-SNAREs Confer Endocytosis
Mol. Biol. Cell, October 1, 2000; 11(10): 3629 - 3643.
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Mol. Biol. CellHome page
D. C. Bassham, A. A. Sanderfoot, V. Kovaleva, H. Zheng, and N. V. Raikhel
AtVPS45 Complex Formation at the trans-Golgi Network
Mol. Biol. Cell, July 1, 2000; 11(7): 2251 - 2265.
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Mol. Biol. CellHome page
M. J. Conboy and M. S. Cyert
Luv1p/Rki1p/Tcs3p/Vps54p, a Yeast Protein That Localizes to the Late Golgi and Early Endosome, Is Required for Normal Vacuolar Morphology.
Mol. Biol. Cell, July 1, 2000; 11(7): 2429 - 2443.
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A. Wiederkehr, S. Avaro, C. Prescianotto-Baschong, R. Haguenauer-Tsapis, and H. Riezman
The F-box Protein Rcy1p Is Involved in Endocytic Membrane Traffic and Recycling Out of an Early Endosome in Saccharomyces cerevisiae
J. Cell Biol., April 17, 2000; 149(2): 397 - 410.
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J. Cell Sci.Home page
H. Panek, E Conibear, J. Bryan, R. Colvin, C. Goshorn, and L. Robinson
Identification of Rgp1p, a novel Golgi recycling factor, as a protein required for efficient localization of yeast casein kinase 1 to the plasma membrane
J. Cell Sci., January 12, 2000; 113(24): 4545 - 4555.
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P. P. Poon, S. F. Nothwehr, R. A. Singer, and G. C. Johnston
The Gcs1 and Age2 ArfGAP proteins provide overlapping essential function for transport from the yeast trans-Golgi network
J. Cell Biol., December 24, 2001; 155(7): 1239 - 1250.
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F. Mallard, B. L. Tang, T. Galli, D. Tenza, A. Saint-Pol, X. Yue, C. Antony, W. Hong, B. Goud, and L. Johannes
Early/recycling endosomes-to-TGN transport involves two SNARE complexes and a Rab6 isoform
J. Cell Biol., February 18, 2002; 156(4): 653 - 664.
[Abstract] [Full Text] [PDF]




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