Identification of Domains Required for Developmentally Regulated SNARE Function in Saccharomyces cerevisiae

Corresponding author: Aaron M. Neiman, Department of Biochemistry and Cell Biology, 332 Life Sciences, State University of New York, Stony Brook, New York 11794-5215., aneiman{at}ms.cc.sunysb.edu (E-mail)

Communicating editor: M. D. ROSE

Saccharomyces cerevisiae cells contain two homologues of the mammalian t-SNARE protein SNAP-25, encoded by the SEC9 and SPO20 genes. Although both gene products participate in post-Golgi vesicle fusion events, they cannot substitute for one another; Sec9p is active primarily in vegetative cells while Spo20p functions only during sporulation. We have investigated the basis for the developmental stage-specific differences in the function of these two proteins. Localization of the other plasma membrane SNARE subunits, Ssop and Sncp, in sporulating cells suggests that these proteins act in conjunction with Spo20p in the formation of the prospore membrane. In vitro binding studies demonstrate that, like Sec9p, Spo20p binds specifically to the t-SNARE Sso1p and, once bound to Sso1p, can complex with the v-SNARE Snc2p. Therefore, Sec9p and Spo20p interact with the same binding partners, but developmental conditions appear to favor the assembly of complexes with Spo20p in sporulating cells. Analysis of chimeric Sec9p/Spo20p molecules indicates that regions in both the SNAP-25 domain and the unique N terminus of Spo20p are required for activity during sporulation. Additionally, the N terminus of Spo20p is inhibitory in vegetative cells. Deletion studies indicate that activation and inhibition are separable functions of the Spo20p N terminus. Our results reveal an additional layer of regulation of the SNARE complex, which is necessary only in sporulating cells.

This article has been cited by other articles:

				Y. Suda, H. Nakanishi, E. M. Mathieson, and A. M. Neiman Alternative Modes of Organellar Segregation during Sporulation in Saccharomyces cerevisiae Eukaryot. Cell, November 1, 2007; 6(11): 2009 - 2017. [Abstract] [Full Text] [PDF]

				M. E. Pablo-Hernando, Y. Arnaiz-Pita, H. Nakanishi, D. Dawson, F. del Rey, A. M. Neiman, and C. R. V. de Aldana Cdc15 Is Required for Spore Morphogenesis Independently of Cdc14 in Saccharomyces cerevisiae Genetics, September 1, 2007; 177(1): 281 - 293. [Abstract] [Full Text] [PDF]

				H. Nakanishi, Y. Suda, and A. M. Neiman Erv14 family cargo receptors are necessary for ER exit during sporulation in Saccharomyces cerevisiae J. Cell Sci., March 1, 2007; 120(5): 908 - 916. [Abstract] [Full Text] [PDF]

				H. Nakanishi, M. Morishita, C. L. Schwartz, A. Coluccio, J. Engebrecht, and A. M. Neiman Phospholipase D and the SNARE Sso1p are necessary for vesicle fusion during sporulation in yeast. J. Cell Sci., April 1, 2006; 119(Pt 7): 1406 - 1415. [Abstract] [Full Text] [PDF]

				A. M. Neiman Ascospore Formation in the Yeast Saccharomyces cerevisiae Microbiol. Mol. Biol. Rev., December 1, 2005; 69(4): 565 - 584. [Abstract] [Full Text] [PDF]

				M. Morishita and J. Engebrecht End3p-Mediated Endocytosis Is Required for Spore Wall Formation in Saccharomyces cerevisiae Genetics, August 1, 2005; 170(4): 1561 - 1574. [Abstract] [Full Text] [PDF]

				A. Coluccio, E. Bogengruber, M. N. Conrad, M. E. Dresser, P. Briza, and A. M. Neiman Morphogenetic Pathway of Spore Wall Assembly in Saccharomyces cerevisiae Eukaryot. Cell, December 1, 2004; 3(6): 1464 - 1475. [Abstract] [Full Text] [PDF]

				A. Coluccio and A. M. Neiman Interspore bridges: a new feature of the Saccharomyces cerevisiae spore wall Microbiology, October 1, 2004; 150(10): 3189 - 3196. [Abstract] [Full Text] [PDF]

				M. E. Nickas, A. E. Diamond, M.-J. Yang, and A. M. Neiman Regulation of Spindle Pole Function by an Intermediary Metabolite Mol. Biol. Cell, June 1, 2004; 15(6): 2606 - 2616. [Abstract] [Full Text] [PDF]

				H. Nakanishi, P. de los Santos, and A. M. Neiman Positive and Negative Regulation of a SNARE Protein by Control of Intracellular Localization Mol. Biol. Cell, April 1, 2004; 15(4): 1802 - 1815. [Abstract] [Full Text] [PDF]

				C. Shimoda Forespore membrane assembly in yeast: coordinating SPBs and membrane trafficking J. Cell Sci., January 22, 2004; 117(3): 389 - 396. [Abstract] [Full Text] [PDF]

				A. Coluccio, M. Malzone, and A. M. Neiman Genetic Evidence of a Role for Membrane Lipid Composition in the Regulation of Soluble NEM-Sensitive Factor Receptor Function in Saccharomyces cerevisiae Genetics, January 1, 2004; 166(1): 89 - 97. [Abstract] [Full Text] [PDF]

				D. Okuzaki, W. Satake, A. Hirata, and H. Nojima Fission yeast meu14+ is required for proper nuclear division and accurate forespore membrane formation during meiosis II J. Cell Sci., July 1, 2003; 116(13): 2721 - 2735. [Abstract] [Full Text] [PDF]

				M. E. Nickas, C. Schwartz, and A. M. Neiman Ady4p and Spo74p Are Components of the Meiotic Spindle Pole Body That Promote Growth of the Prospore Membrane in Saccharomyces cerevisiae Eukaryot. Cell, June 1, 2003; 2(3): 431 - 445. [Abstract] [Full Text] [PDF]

				M. E. Nickas and A. M. Neiman Ady3p Links Spindle Pole Body Function to Spore Wall Synthesis in Saccharomyces cerevisiae Genetics, April 1, 2002; 160(4): 1439 - 1450. [Abstract] [Full Text] [PDF]

				T. Nakamura, M. Nakamura-Kubo, A. Hirata, and C. Shimoda The Schizosaccharomyces pombe spo3+ Gene Is Required for Assembly of the Forespore Membrane and Genetically Interacts with psy1+-encoding Syntaxin-like Protein Mol. Biol. Cell, December 1, 2001; 12(12): 3955 - 3972. [Abstract] [Full Text] [PDF]

				B. K. Bajgier, M. Malzone, M. Nickas, and A. M. Neiman SPO21 Is Required for Meiosis-specific Modification of the Spindle Pole Body in Yeast Mol. Biol. Cell, June 1, 2001; 12(6): 1611 - 1621. [Abstract] [Full Text] [PDF]

				A. A. Sanderfoot, F. F. Assaad, and N. V. Raikhel The Arabidopsis Genome. An Abundance of Soluble N-Ethylmaleimide-Sensitive Factor Adaptor Protein Receptors Plant Physiology, December 1, 2000; 124(4): 1558 - 1569. [Abstract] [Full Text]

				L. C. Gonzalez Jr., W. I. Weis, and R. H. Scheller A Novel SNARE N-terminal Domain Revealed by the Crystal Structure of Sec22b J. Biol. Chem., June 22, 2001; 276(26): 24203 - 24211. [Abstract] [Full Text] [PDF]

				H. Tachikawa, A. Bloecher, K. Tatchell, and A. M. Neiman A Gip1p-Glc7p phosphatase complex regulates septin organization and spore wall formation J. Cell Biol., November 26, 2001; 155(5): 797 - 808. [Abstract] [Full Text] [PDF]

				M. Heese, X. Gansel, L. Sticher, P. Wick, M. Grebe, F. Granier, and G. Jurgens Functional characterization of the KNOLLE-interacting t-SNARE AtSNAP33 and its role in plant cytokinesis J. Cell Biol., October 15, 2001; 155(2): 239 - 250. [Abstract] [Full Text] [PDF]