Pep12p is a Multifunctional Yeast Syntaxin that Controls Entry of Biosynthetic, Endocytic and Retrograde Traffic into the Prevacuolar Compartment

Delivery of proteins to the vacuole of the yeast Saccharomyces cerevisiae requires the function of the endosomal syntaxin, Pep12p. Many vacuolar proteins, such as the soluble vacuolar hydrolase, carboxypeptidase Y (CPY), traverse the prevacuolar compartment (PVC) en route to the vacuole. Here we show that deletion of the carboxy‐terminal transmembrane domain of Pep12p results in a temperature‐conditional block in transport of CPY to the PVC. The PVC also receives traffic from the early endosome and the vacuole, and mutation in PEP12 also blocks these other trafficking pathways into the PVC. Therefore, Pep12p is a multifunctional syntaxin that is required for all known trafficking pathways into the yeast PVC. Finally, we found that the internalized pheromone receptor, Ste3p, can cycle out of the PVC in a VPS27‐independent fashion.

[1]  S. Emr,et al.  Mutations in the VPS45 gene, a SEC1 homologue, result in vacuolar protein sorting defects and accumulation of membrane vesicles. , 1994, Journal of cell science.

[2]  Reinhard Jahn,et al.  Vesicle fusion from yeast to man , 1994, Nature.

[3]  S. Emr,et al.  Fab1p PtdIns(3)P 5-Kinase Function Essential for Protein Sorting in the Multivesicular Body , 1998, Cell.

[4]  T. Stevens,et al.  Vps10p cycles between the late-Golgi and prevacuolar compartments in its function as the sorting receptor for multiple yeast vacuolar hydrolases , 1996, The Journal of cell biology.

[5]  C. J. Roberts,et al.  Membrane protein sorting in the yeast secretory pathway: evidence that the vacuole may be the default compartment , 1992, The Journal of cell biology.

[6]  T. Stevens,et al.  Vacuole Biogenesis in Saccharomyces cerevisiae: Protein Transport Pathways to the Yeast Vacuole , 1998, Microbiology and Molecular Biology Reviews.

[7]  G. Sprague,,et al.  Cis- and trans-acting functions required for endocytosis of the yeast pheromone receptors , 1993, The Journal of cell biology.

[8]  T. Stevens,et al.  The Yeast Endosomal t‐SNARE, Pep12p, Functions in the Absence of its Transmembrane Domain , 2000, Traffic.

[9]  T. Stevens,et al.  The Yeast v-SNARE Vti1p Mediates Two Vesicle Transport Pathways through Interactions with the t-SNAREs Sed5p and Pep12p , 1997, The Journal of cell biology.

[10]  S. Emr,et al.  The cytoplasmic tail domain of the vacuolar protein sorting receptor Vps10p and a subset of VPS gene products regulate receptor stability, function, and localization. , 1995, Molecular biology of the cell.

[11]  T. Stevens,et al.  An MBoC Favorite: Morphological classification of the yeast vacuolar protein-sorting mutants: evidence for a prevacuolar compartment in class E vps mutants , 1992, Molecular biology of the cell.

[12]  P. Philippsen,et al.  New heterologous modules for classical or PCR‐based gene disruptions in Saccharomyces cerevisiae , 1994, Yeast.

[13]  S. Emr,et al.  A Multispecificity Syntaxin Homologue, Vam3p, Essential for Autophagic and Biosynthetic Protein Transport to the Vacuole , 1997, The Journal of cell biology.

[14]  T. Stevens,et al.  VPS27 controls vacuolar and endocytic traffic through a prevacuolar compartment in Saccharomyces cerevisiae , 1995, The Journal of cell biology.

[15]  W. B. Snyder,et al.  Novel Golgi to vacuole delivery pathway in yeast: identification of a sorting determinant and required transport component , 1997, The EMBO journal.

[16]  S. Emr,et al.  VPS21 encodes a rab5‐like GTP binding protein that is required for the sorting of yeast vacuolar proteins. , 1994, The EMBO journal.

[17]  C. J. Roberts,et al.  Membrane protein retention in the yeast Golgi apparatus: dipeptidyl aminopeptidase A is retained by a cytoplasmic signal containing aromatic residues , 1993, The Journal of cell biology.

[18]  T. Stevens,et al.  Golgi and vacuolar membrane proteins reach the vacuole in vps1 mutant yeast cells via the plasma membrane , 1995, The Journal of cell biology.

[19]  T. Stevens,et al.  Protein sorting in yeast: Mutants defective in vacuole biogenesis mislocalize vacuolar proteins into the late secretory pathway , 1986, Cell.

[20]  S. Emr,et al.  Novel syntaxin homologue, Pep12p, required for the sorting of lumenal hydrolases to the lysosome-like vacuole in yeast. , 1996, Molecular biology of the cell.

[21]  T. Stevens,et al.  Yeast Vps45p is a Sec1p-like protein required for the consumption of vacuole-targeted, post-Golgi transport vesicles. , 1994, European journal of cell biology.

[22]  S. Emr,et al.  The sorting receptor for yeast vacuolar carboxypeptidase Y is encoded by the VPS10 gene , 1994, Cell.

[23]  T. Stevens,et al.  The Membrane Protein Alkaline Phosphatase Is Delivered to the Vacuole by a Route That Is Distinct from the VPS-dependent Pathway , 1997, The Journal of cell biology.

[24]  Paul Tempst,et al.  SNAP receptors implicated in vesicle targeting and fusion , 1993, Nature.

[25]  S. Emr,et al.  The VPS16 gene product associates with a sedimentable protein complex and is essential for vacuolar protein sorting in yeast. , 1993, The Journal of biological chemistry.

[26]  T. Stevens,et al.  The Saccharomyces cerevisiae v-SNARE Vti1p is required for multiple membrane transport pathways to the vacuole. , 1999, Molecular biology of the cell.

[27]  S. Lemmon,et al.  The Yeast Adaptor Protein Complex, AP-3, Is Essential for the Efficient Delivery of Alkaline Phosphatase by the Alternate Pathway to the Vacuole , 1997, The Journal of cell biology.

[28]  T. Stevens,et al.  VPS21 controls entry of endocytosed and biosynthetic proteins into the yeast prevacuolar compartment. , 2000, Molecular biology of the cell.

[29]  T. Stevens,et al.  Retrograde Traffic Out of the Yeast Vacuole to the TGN Occurs via the Prevacuolar/Endosomal Compartment , 1998, The Journal of cell biology.

[30]  Natalie L. Catlett,et al.  Vac7p, a novel vacuolar protein, is required for normal vacuole inheritance and morphology , 1997, Molecular and cellular biology.

[31]  T. Stevens,et al.  Traffic into the prevacuolar/endosomal compartment of Saccharomyces cerevisiae: a VPS45-dependent intracellular route and a VPS45-independent, endocytic route. , 1998, European journal of cell biology.

[32]  P. Philippsen,et al.  Role of three rab5-like GTPases, Ypt51p, Ypt52p, and Ypt53p, in the endocytic and vacuolar protein sorting pathways of yeast , 1994, The Journal of cell biology.

[33]  T. Stevens,et al.  Two Separate Signals Act Independently to Localize a Yeast Late Golgi Membrane Protein through a Combination of Retrieval and Retention , 1997, The Journal of cell biology.

[34]  J. Rothman,et al.  Mechanisms of intracellular protein transport , 1994, Nature.

[35]  C. J. Roberts,et al.  Methods for studying the yeast vacuole. , 1991, Methods in enzymology.

[36]  J. McNew,et al.  Characterization of a novel yeast SNARE protein implicated in Golgi retrograde traffic. , 1997, Molecular biology of the cell.