The Journal of Cell Biology, Vol 135, 597-610, Copyright © 1996 by The Rockefeller University Press

ARTICLES

Differential trafficking and timed localization of two chitin synthase proteins, Chs2p and Chs3p [published erratum appears in J Cell Biol 1996 Dec;135(6 Pt 2):1925]

JS Chuang and RW Schekman
Department of Molecular and Cell Biology, University of California, Berkeley 94720, USA.

The deposition of the polysaccharide chitin in the Saccharomyces cerevisiae cell wall is temporally and spatially regulated. Chitin synthase III (Chs3p) synthesizes a ring of chitin at the onset of bud emergence, marking the base of the incipient bud. At the end of mitosis, chitin synthase II (Chs2p) deposits a disk of chitin in the mother-bud neck, forming the primary division septum. Using indirect immunofluorescence microscopy, we have found that these two integral membrane proteins localize to the mother-bud neck at distinct times during the cell cycle. Chs2p is found at the neck at the end of mitosis, whereas Chs3p localizes to a ring on the surface of cells about to undergo bud emergence and in the mother-bud neck of small- budded cells. Cell synchronization and pulse-chase experiments suggest that the timing of Chs2p localization results from cell cycle-specific synthesis coupled to rapid degradation. Chs2p degradation depends on the vacuolar protease encoded by PEP4, indicating that Chs2p is destroyed in the vacuole. Temperature-sensitive mutations that block either the late secretory pathway (sec1-1) or the internalization step of endocytosis (end4-1) also prevent Chs2p degradation. In contrast, Chs3p is synthesized constitutively and is metabolically stable, indicating that Chs2p and Chs3p are subject to different modes of regulation. Differential centrifugation experiments show that a significant proportion of Chs3p resides in an internal compartment that may correspond to a vesicular species called the chitosome (Leal- Morales, C.A., C.E. Bracker, and S. Bartnicki-Garcia. 1988, Proc. Natl. Acad. Sci. USA. 85:8516-8520; Flores Martinez, A., and J. Schwencke. 1988. Biochim. Biophys. Acta. 946:328-336). Fractionation of membranes prepared from mutants defective in internalization (end3-1 and end4-1) indicate that the Chs3p-containing vesicles are endocytically derived. Collectively, these data suggest that the trafficking of Chs2p and Chs3p diverges after endocytosis; Chs3p is not delivered to the vacuole, but instead may be recycled.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Collette, J. R., Chi, R. J., Boettner, D. R., Fernandez-Golbano, I. M., Plemel, R., Merz, A. J., Geli, M. I., Traub, L. M., Lemmon, S. K. (2009). Clathrin Functions in the Absence of the Terminal Domain Binding Site for Adaptor-associated Clathrin-Box Motifs. Mol. Biol. Cell 20: 3401-3413 [Abstract] [Full Text]  
• Nishihama, R., Schreiter, J. H., Onishi, M., Vallen, E. A., Hanna, J., Moravcevic, K., Lippincott, M. F., Han, H., Lemmon, M. A., Pringle, J. R., Bi, E. (2009). Role of Inn1 and its interactions with Hof1 and Cyk3 in promoting cleavage furrow and septum formation in S. cerevisiae. JCB 185: 995-1012 [Abstract] [Full Text]  
• Wiederhold, E., Gandhi, T., Permentier, H. P., Breitling, R., Poolman, B., Slotboom, D. J. (2009). The Yeast Vacuolar Membrane Proteome. Mol. Cell. Proteomics 8: 380-392 [Abstract] [Full Text]  
• Larson, J. R., Bharucha, J. P., Ceaser, S., Salamon, J., Richardson, C. J., Rivera, S. M., Tatchell, K. (2008). Protein Phosphatase Type 1 Directs Chitin Synthesis at the Bud Neck in Saccharomyces cerevisiae. Mol. Biol. Cell 19: 3040-3051 [Abstract] [Full Text]  
• Mesecke, N., Spang, A., Deponte, M., Herrmann, J. M. (2008). A Novel Group of Glutaredoxins in the cis-Golgi Critical for Oxidative Stress Resistance. Mol. Biol. Cell 19: 2673-2680 [Abstract] [Full Text]  
• Riquelme, M., Bartnicki-Garcia, S., Gonzalez-Prieto, J. M., Sanchez-Leon, E., Verdin-Ramos, J. A., Beltran-Aguilar, A., Freitag, M. (2007). Spitzenkorper Localization and Intracellular Traffic of Green Fluorescent Protein-Labeled CHS-3 and CHS-6 Chitin Synthases in Living Hyphae of Neurospora crassa. Eukaryot Cell 6: 1853-1864 [Abstract] [Full Text]  
• Reyes, A., Sanz, M., Duran, A., Roncero, C. (2007). Chitin synthase III requires Chs4p-dependent translocation of Chs3p into the plasma membrane. J. Cell Sci. 120: 1998-2009 [Abstract] [Full Text]  
• Copic, A., Starr, T. L., Schekman, R. (2007). Ent3p and Ent5p Exhibit Cargo-specific Functions in Trafficking Proteins between the Trans-Golgi Network and the Endosomes in Yeast. Mol. Biol. Cell 18: 1803-1815 [Abstract] [Full Text]  
• Duncan, M. C., Ho, D. G., Huang, J., Jung, M. E., Payne, G. S. (2007). Composite synthetic lethal identification of membrane traffic inhibitors. Proc. Natl. Acad. Sci. USA 104: 6235-6240 [Abstract] [Full Text]  
• Cameroni, E., De Virgilio, C., Deloche, O. (2006). Phosphatidylinositol 4-Phosphate Is Required for Translation Initiation in Saccharomyces cerevisiae. J. Biol. Chem. 281: 38139-38149 [Abstract] [Full Text]  
• Sanchatjate, S., Schekman, R. (2006). Chs5/6 Complex: A Multiprotein Complex That Interacts with and Conveys Chitin Synthase III from the Trans-Golgi Network to the Cell Surface. Mol. Biol. Cell 17: 4157-4166 [Abstract] [Full Text]  
• Moseley, J. B., Goode, B. L. (2006). The Yeast Actin Cytoskeleton: from Cellular Function to Biochemical Mechanism. Microbiol. Mol. Biol. Rev. 70: 605-645 [Abstract] [Full Text]  
• Osterberg, M., Kim, H., Warringer, J., Melen, K., Blomberg, A., von Heijne, G. (2006). Phenotypic effects of membrane protein overexpression in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 103: 11148-11153 [Abstract] [Full Text]  
• Zhang, G., Kashimshetty, R., Ng, K. E., Tan, H. B., Yeong, F. M. (2006). Exit from mitosis triggers Chs2p transport from the endoplasmic reticulum to mother-daughter neck via the secretory pathway in budding yeast. JCB 174: 207-220 [Abstract] [Full Text]  
• Smits, G. J., Schenkman, L. R., Brul, S., Pringle, J. R., Klis, F. M. (2006). Role of Cell Cycle-regulated Expression in the Localized Incorporation of Cell Wall Proteins in Yeast. Mol. Biol. Cell 17: 3267-3280 [Abstract] [Full Text]  
• Lesage, G., Bussey, H. (2006). Cell Wall Assembly in Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 70: 317-343 [Abstract] [Full Text]  
• Cai, H., Zhang, Y., Pypaert, M., Walker, L., Ferro-Novick, S. (2005). Mutants in trs120 disrupt traffic from the early endosome to the late Golgi. JCB 171: 823-833 [Abstract] [Full Text]  
• Ichinomiya, M., Yamada, E., Yamashita, S., Ohta, A., Horiuchi, H. (2005). Class I and Class II Chitin Synthases Are Involved in Septum Formation in the Filamentous Fungus Aspergillus nidulans. Eukaryot Cell 4: 1125-1136 [Abstract] [Full Text]  
• VerPlank, L., Li, R. (2005). Cell Cycle-regulated Trafficking of Chs2 Controls Actomyosin Ring Stability during Cytokinesis. Mol. Biol. Cell 16: 2529-2543 [Abstract] [Full Text]  
• Malkus, P., Graham, L. A., Stevens, T. H., Schekman, R. (2004). Role of Vma21p in Assembly and Transport of the Yeast Vacuolar ATPase. Mol. Biol. Cell 15: 5075-5091 [Abstract] [Full Text]  
• Kipnis, P., Thomas, N., Ovalle, R., Lipke, P. N. (2004). The ER-Golgi v-SNARE Bet1p is required for cross-linking {alpha}-agglutinin to the cell wall in yeast. Microbiology 150: 3219-3228 [Abstract] [Full Text]  
• Selvaggini, S., Munro, C. A., Paschoud, S., Sanglard, D., Gow, N. A. R. (2004). Independent regulation of chitin synthase and chitinase activity in Candida albicans and Saccharomyces cerevisiae. Microbiology 150: 921-928 [Abstract] [Full Text]  
• de la Cruz, J., Lacombe, T., Deloche, O., Linder, P., Kressler, D. (2004). The Putative RNA Helicase Dbp6p Functionally Interacts With Rpl3p, Nop8p and the Novel trans-acting Factor Rsa3p During Biogenesis of 60S Ribosomal Subunits in Saccharomyces cerevisiae. Genetics 166: 1687-1699 [Abstract] [Full Text]  
• Hwa Lim, H., Yeong, F. M., Surana, U. (2003). Inactivation of Mitotic Kinase Triggers Translocation of MEN Components to Mother-Daughter Neck in Yeast. Mol. Biol. Cell 14: 4734-4743 [Abstract] [Full Text]  
• Bulik, D. A., Olczak, M., Lucero, H. A., Osmond, B. C., Robbins, P. W., Specht, C. A. (2003). Chitin Synthesis in Saccharomyces cerevisiae in Response to Supplementation of Growth Medium with Glucosamine and Cell Wall Stress. Eukaryot Cell 2: 886-900 [Abstract] [Full Text]  
• Valdivia, R. H., Schekman, R. (2003). The yeasts Rho1p and Pkc1p regulate the transport of chitin synthase III (Chs3p) from internal stores to the plasma membrane. Proc. Natl. Acad. Sci. USA 100: 10287-10292 [Abstract] [Full Text]  
• Sandmann, T., Herrmann, J. M., Dengjel, J., Schwarz, H., Spang, A. (2003). Suppression of Coatomer Mutants by a New Protein Family with COPI and COPII Binding Motifs in Saccharomyces cerevisiae. Mol. Biol. Cell 14: 3097-3113 [Abstract] [Full Text]  
• Misu, K., Fujimura-Kamada, K., Ueda, T., Nakano, A., Katoh, H., Tanaka, K. (2003). Cdc50p, a Conserved Endosomal Membrane Protein, Controls Polarized Growth in Saccharomyces cerevisiae. Mol. Biol. Cell 14: 730-747 [Abstract] [Full Text]  
• Kozubowski, L., Panek, H., Rosenthal, A., Bloecher, A., DeMarini, D. J., Tatchell, K. (2003). A Bni4-Glc7 Phosphatase Complex That Recruits Chitin Synthase to the Site of Bud Emergence. Mol. Biol. Cell 14: 26-39 [Abstract] [Full Text]  
• Deloche, O., Schekman, R. W. (2002). Vps10p Cycles between the TGN and the Late Endosome via the Plasma Membrane in Clathrin Mutants. Mol. Biol. Cell 13: 4296-4307 [Abstract] [Full Text]  
• Cortes, J. C. G., Ishiguro, J., Duran, A., Ribas, J. C. (2002). Localization of the (1,3){beta}-D-glucan synthase catalytic subunit homologue Bgs1p/Cps1p from fission yeast suggests that it is involved in septation, polarized growth, mating, spore wall formation and spore germination. J. Cell Sci. 115: 4081-4096 [Abstract] [Full Text]  
• Hua, Z., Fatheddin, P., Graham, T. R. (2002). An Essential Subfamily of Drs2p-related P-Type ATPases Is Required for Protein Trafficking between Golgi Complex and Endosomal/Vacuolar System. Mol. Biol. Cell 13: 3162-3177 [Abstract] [Full Text]  
• Roh, D.-H., Bowers, B., Schmidt, M., Cabib, E. (2002). The Septation Apparatus, an Autonomous System in Budding Yeast. Mol. Biol. Cell 13: 2747-2759 [Abstract] [Full Text]  
• Rodriguez-Pena, J. M., Rodriguez, C., Alvarez, A., Nombela, C., Arroyo, J. (2002). Mechanisms for targeting of the Saccharomyces cerevisiae GPI-anchored cell wall protein Crh2p to polarised growth sites. J. Cell Sci. 115: 2549-2558 [Abstract] [Full Text]  
• Schmidt, M., Bowers, B., Varma, A., Roh, D.-H., Cabib, E. (2002). In budding yeast, contraction of the actomyosin ring and formation of the primary septum at cytokinesis depend on each other. J. Cell Sci. 115: 293-302 [Abstract] [Full Text]  
• Dupre, S., Haguenauer-Tsapis, R. (2001). Deubiquitination Step in the Endocytic Pathway of Yeast Plasma Membrane Proteins: Crucial Role of Doa4p Ubiquitin Isopeptidase. Mol. Cell. Biol. 21: 4482-4494 [Abstract] [Full Text]  
• Smits, G. J., van den Ende, H., Klis, F. M. (2001). Differential regulation of cell wall biogenesis during growth and development in yeast. Microbiology 147: 781-794 [Full Text]  
• Spang, A., Herrmann, J. M., Hamamoto, S., Schekman, R. (2001). The ADP Ribosylation Factor-Nucleotide Exchange Factors Gea1p and Gea2p Have Overlapping, but Not Redundant Functions in Retrograde Transport from the Golgi to the Endoplasmic Reticulum. Mol. Biol. Cell 12: 1035-1045 [Abstract] [Full Text]  
• Deloche, O., Yeung, B. G., Payne, G. S., Schekman, R. (2001). Vps10p Transport from the trans-Golgi Network to the Endosome Is Mediated by Clathrin-coated Vesicles. Mol. Biol. Cell 12: 475-485 [Abstract] [Full Text]  
• Chen, L., Davis, N. G. (2000). Recycling of the Yeast a-Factor Receptor. JCB 151: 731-738 [Abstract] [Full Text]  
• Valdivieso, M-H., Ferrario, L., Vai, M., Duran, A., Popolo, L. (2000). Chitin Synthesis in a gas1 Mutant of Saccharomyces cerevisiae. J. Bacteriol. 182: 4752-4757 [Abstract] [Full Text]  
• García-Rodriguez, L. J., Durán, A., Roncero, C. (2000). Calcofluor Antifungal Action Depends on Chitin and a Functional High-Osmolarity Glycerol Response (HOG) Pathway: Evidence for a Physiological Role of the Saccharomyces cerevisiae HOG Pathway under Noninducing Conditions. J. Bacteriol. 182: 2428-2437 [Abstract] [Full Text]  
• Wiederkehr, A., Avaro, S., Prescianotto-Baschong, C., Haguenauer-Tsapis, R., Riezman, H. (2000). The F-Box Protein Rcy1p Is Involved in Endocytic Membrane Traffic and Recycling Out of an Early Endosome in Saccharomyces cerevisiae. JCB 149: 397-410 [Abstract] [Full Text]  
• Wang, Z., Szaniszlo, P. J. (2000). WdCHS3, a Gene That Encodes a Class III Chitin Synthase in Wangiella (Exophiala) dermatitidis, Is Expressed Differentially under Stress Conditions. J. Bacteriol. 182: 874-881 [Abstract] [Full Text]  
• Ono, N., Yabe, T., Sudoh, M., Nakajima, T., Yamada-Okabe, T., Arisawa, M., Yamada-Okabe, H. (2000). The yeast Chs4 protein stimulates the trypsin-sensitive activity of chitin synthase 3 through an apparent protein-protein interaction. Microbiology 146: 385-391 [Abstract] [Full Text]  
• Santos, B., Snyder, M. (2000). Sbe2p and Sbe22p, Two Homologous Golgi Proteins Involved in Yeast Cell Wall Formation. Mol. Biol. Cell 11: 435-452 [Abstract] [Full Text]  
• Luo, W.-j., Chang, A. (2000). An Endosome-to-Plasma Membrane Pathway Involved in Trafficking of a Mutant Plasma Membrane ATPase in Yeast. Mol. Biol. Cell 11: 579-592 [Abstract] [Full Text]  
• Vallen, E. A., Caviston, J., Bi, E. (2000). Roles of Hof1p, Bni1p, Bnr1p, and Myo1p in Cytokinesis in Saccharomyces cerevisiae. Mol. Biol. Cell 11: 593-611 [Abstract] [Full Text]  
• Pruyne, D, Bretscher, A (2000). Polarization of cell growth in yeast. J. Cell Sci. 113: 571-585 [Abstract]  
• Trilla, J. A., Duran, A., Roncero, C. (1999). Chs7p, a New Protein Involved in the Control of Protein Export from the Endoplasmic Reticulum that Is Specifically Engaged in the Regulation of Chitin Synthesis in Saccharomyces cerevisiae. JCB 145: 1153-1163 [Abstract] [Full Text]  
• Reck-Peterson, S. L., Novick, P. J., Mooseker, M. S. (1999). The Tail of a Yeast Class V Myosin, Myo2p, Functions as a Localization Domain. Mol. Biol. Cell 10: 1001-1017 [Abstract] [Full Text]  
• Robinson, L. C., Bradley, C., Bryan, J. D., Jerome, A., Kweon, Y., Panek, H. R. (1999). The Yck2 Yeast Casein Kinase 1 Isoform Shows Cell Cycle-specific Localization to Sites of Polarized Growth and Is Required for Proper Septin Organization. Mol. Biol. Cell 10: 1077-1092 [Abstract] [Full Text]  
• Holthuis, J. C. M., Nichols, B. J., Pelham, H. R. B. (1998). The Syntaxin Tlg1p Mediates Trafficking of Chitin Synthase III to Polarized Growth Sites in Yeast. Mol. Biol. Cell 9: 3383-3397 [Abstract] [Full Text]  
• Ziman, M., Chuang, J. S., Tsung, M., Hamamoto, S., Schekman, R. (1998). Chs6p-dependent Anterograde Transport of Chs3p from the Chitosome to the Plasma Membrane in Saccharomyces cerevisiae. Mol. Biol. Cell 9: 1565-1576 [Abstract] [Full Text]  
• DeMarini, D. J., Adams, A. E.M., Fares, H., Virgilio, C. D., Valle, G., Chuang, J. S., Pringle, J. R. (1997). A Septin-based Hierarchy of Proteins Required for Localized Deposition of Chitin in the Saccharomyces cerevisiae Cell Wall. JCB 139: 75-93 [Abstract] [Full Text]  
• Jedd, G., Mulholland, J., Segev, N. (1997). Two New Ypt GTPases Are Required for Exit From the Yeast trans-Golgi Compartment. JCB 137: 563-580 [Abstract] [Full Text]  
• Cabib, E., Roh, D.-H., Schmidt, M., Crotti, L. B., Varma, A. (2001). The Yeast Cell Wall and Septum as Paradigms of Cell Growth and Morphogenesis. J. Biol. Chem. 276: 19679-19682 [Full Text]  
• Todorow, Z., Spang, A., Carmack, E., Yates, J., Schekman, R. (2000). Active recycling of yeast Golgi mannosyltransferase complexes through the endoplasmic reticulum. Proc. Natl. Acad. Sci. USA 97: 13643-13648 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents