J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/01/95/16 $2.00
Volume 136, Number 1, January 13, 1997 95-110

Targeting of Chitin Synthase 3 to Polarized Growth Sites in Yeast Requires Chs5p and Myo2p

Beatriz Santos, and Michael Snyder

Department of Biology, Yale University, New Haven, Connecticut 06520-8103

Chitin is an essential structural component of the yeast cell wall whose deposition is regulated throughout the yeast life cycle. The temporal and spatial regulation of chitin synthesis was investigated during vegetative growth and mating of Saccharomyces cerevisiae by localization of the putative catalytic subunit of chitin synthase III, Chs3p, and its regulator, Chs5p. Immunolocalization of epitope-tagged Chs3p revealed a novel localization pattern that is cell cycledependent. Chs3p is polarized as a diffuse ring at the incipient bud site and at the neck between the mother and bud in small-budded cells; it is not found at the neck in large-budded cells containing a single nucleus. In large-budded cells undergoing cytokinesis, it reappears as a ring at the neck. In cells responding to mating pheromone, Chs3p is found throughout the projection. The appearance of Chs3p at cortical sites correlates with times that chitin synthesis is expected to occur. In addition to its localization at the incipient bud site and neck, Chs3p is also found in cytoplasmic patches in cells at different stages of the cell cycle. Epitope-tagged Chs5p also localizes to cytoplasmic patches; these patches contain Kex2p, a late Golgi-associated enzyme. Unlike Chs3p, Chs5p does not accumulate at the incipient bud site or neck. Nearly all Chs3p patches contain Chs5p, whereas some Chs5p patches lack detectable Chs3p. In the absence of Chs5p, Chs3p localizes in cytoplasmic patches, but it is no longer found at the neck or the incipient bud site, indicating that Chs5p is required for the polarization of Chs3p. Furthermore, Chs5p localization is not affected either by temperature shift or by the myo2-66 mutation, however, Chs3p polarization is affected by temperature shift and myo2-66. We suggest a model in which Chs3p polarization to cortical sites in yeast is dependent on both Chs5p and the actin cytoskeleton/Myo2p.

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• Rossanese, O. W., Reinke, C. A., Bevis, B. J., Hammond, A. T., Sears, I. B., O'Connor, J., Glick, B. S. (2001). A Role for Actin, Cdc1p, and Myo2p in the Inheritance of Late Golgi Elements in Saccharomyces cerevisiae. J. Cell Biol. 153: 47-62 [Abstract] [Full Text]  
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• 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]  
• Catlett, N. L., Duex, J. E., Tang, F., Weisman, L. S. (2000). Two Distinct Regions in a Yeast Myosin-V Tail Domain Are Required for the Movement of Different Cargoes. J. Cell Biol. 150: 513-526 [Abstract] [Full Text]  
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• 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]  
• Pruyne, D, Bretscher, A (2000). Polarization of cell growth in yeast. J. Cell Sci. 113: 571-585 [Abstract]  
• Schott, D., Ho, J., Pruyne, D., Bretscher, A. (1999). The COOH-Terminal Domain of Myo2p, a Yeast Myosin V, Has a Direct Role in Secretory Vesicle Targeting. J. Cell Biol. 147: 791-808 [Abstract] [Full Text]  
• Osmond, B. C., Specht, C. A., Robbins, P. W. (1999). Chitin synthase III: Synthetic lethal mutants and "stress related" chitin synthesis that bypasses the CSD3/CHS6 localization pathway. Proc. Natl. Acad. Sci. USA 96: 11206-11210 [Abstract] [Full Text]  
• Horiuchi, H., Fujiwara, M., Yamashita, S., Ohta, A., Takagi, M. (1999). Proliferation of Intrahyphal Hyphae Caused by Disruption of csmA, Which Encodes a Class V Chitin Synthase with a Myosin Motor-Like Domain in Aspergillus nidulans. J. Bacteriol. 181: 3721-3729 [Abstract] [Full Text]  
• 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. J. Cell Biol. 145: 1153-1163 [Abstract] [Full Text]  
• Lustgarten, V., Gerst, J. E. (1999). Yeast VSM1 Encodes a v-SNARE Binding Protein That May Act as a Negative Regulator of Constitutive Exocytosis. Mol. Cell. Biol. 19: 4480-4494 [Abstract] [Full Text]  
• Heimann, K., Percival, J. M., Weinberger, R., Gunning, P., Stow, J. L. (1999). Specific Isoforms of Actin-binding Proteins on Distinct Populations of Golgi-derived Vesicles. J. Biol. Chem. 274: 10743-10750 [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]  
• Catlett, N. L., Weisman, L. S. (1998). The terminal tail region of a yeast myosin-V mediates its attachment to vacuole membranes and sites of polarized growth. Proc. Natl. Acad. Sci. USA 95: 14799-14804 [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]  
• Longtine, M. S., Fares, H., Pringle, J. R. (1998). Role of the Yeast Gin4p Protein Kinase in Septin Assembly and the Relationship between Septin Assembly and Septin Function. J. Cell Biol. 143: 719-736 [Abstract] [Full Text]  
• Sheu, Y.-J., Santos, B., Fortin, N., Costigan, C., Snyder, M. (1998). Spa2p Interacts with Cell Polarity Proteins and Signaling Components Involved in Yeast Cell Morphogenesis. Mol. Cell. Biol. 18: 4053-4069 [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]  
• Tabb, J., Molyneaux, B., Cohen, D., Kuznetsov, S., Langford, G. (1998). Transport of ER vesicles on actin filaments in neurons by myosin V. J. Cell Sci. 111: 3221-3234 [Abstract]  
• Roemer, T, Vallier, L, Sheu, Y., Snyder, M (1998). The Spa2-related protein, Sph1p, is important for polarized growth in yeast. J. Cell Sci. 111: 479-494 [Abstract]  
• 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. J. Cell Biol. 139: 75-93 [Abstract] [Full Text]  
• Manning, B. D., Padmanabha, R., Snyder, M. (1997). The Rho-GEF Rom2p Localizes to Sites of Polarized Cell Growth and Participates in Cytoskeletal Functions in Saccharomyces cerevisiae. Mol. Biol. Cell 8: 1829-1844 [Abstract] [Full Text]  
• Nascimento, A. A.C., Amaral, R. G., Bizario, J. C.S., Larson, R. E., Espreafico, E. M. (1997). Subcellular Localization of Myosin-V in the B16 Melanoma Cells, a Wild-type Cell Line for the dilute Gene. Mol. Biol. Cell 8: 1971-1988 [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]  

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