J. Cell Biol.
© The Rockefeller University Press
0021-9525/97/01/251/19 $2.00
Volume 136, Number 2, January 27, 1997 251-269

Biogenesis of the Saccharomyces cerevisiae Mating Pheromone a-Factor

Peng Chen, Stephanie K. Sapperstein, Jonathan D. Choi, and Susan Michaelis

Department of Cell Biology and Anatomy, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

The Saccharomyces cerevisiae mating pheromone a-factor is a prenylated and carboxyl methylated extracellular peptide signaling molecule. Biogenesis of the a-factor precursor proceeds via a distinctive multistep pathway that involves COOH-terminal modification, NH₂-terminal proteolysis, and a nonclassical export mechanism. In this study, we examine the formation and fate of a-factor biosynthetic intermediates to more precisely define the events that occur during a-factor biogenesis. We have identified four distinct a-factor biosynthetic intermediates (P0, P1, P2, and M) by metabolic labeling, immunoprecipitation, and SDSPAGE. We determined the biochemical composition of each by defining their NH₂-terminal amino acid and COOH-terminal modification status. Unexpectedly, we discovered that not one, but two NH₂-terminal cleavage steps occur during the biogenesis of a-factor. In addition, we have shown that COOH-terminal prenylation is required for the NH₂-terminal processing of a-factor and that all the prenylated a-factor intermediates (P1, P2, and M) are membrane bound, suggesting that many steps of a-factor biogenesis occur in association with membranes. We also observed that although the biogenesis of a-factor is a rapid process, it is inherently inefficient, perhaps reflecting the potential for regulation. Previous studies have identified gene products that participate in the COOH-terminal modification (Ram1p, Ram2p, Ste14p), NH₂-terminal processing (Ste24p, Axl1p), and export (Ste6p) of a-factor. The intermediates defined in the present study are discussed in the context of these biogenesis components to formulate an overall model for the pathway of a-factor biogenesis.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

• Muller, H., Hennequin, C., Gallaud, J., Dujon, B., Fairhead, C. (2008). The Asexual Yeast Candida glabrata Maintains Distinct a and {alpha} Haploid Mating Types. Eukaryot Cell 7: 848-858 [Abstract] [Full Text]  
• Hoff, B., Poggeler, S., Kuck, U. (2008). Eighty Years after Its Discovery, Fleming's Penicillium Strain Discloses the Secret of Its Sex. Eukaryot Cell 7: 465-470 [Abstract] [Full Text]  
• Yi, S., Sahni, N., Daniels, K. J., Pujol, C., Srikantha, T., Soll, D. R. (2008). The Same Receptor, G Protein, and Mitogen-activated Protein Kinase Pathway Activate Different Downstream Regulators in the Alternative White and Opaque Pheromone Responses of Candida albicans. Mol. Biol. Cell 19: 957-970 [Abstract] [Full Text]  
• Dignard, D., El-Naggar, A. L., Logue, M. E., Butler, G., Whiteway, M. (2007). Identification and Characterization of MFA1, the Gene Encoding Candida albicans a-Factor Pheromone. Eukaryot Cell 6: 487-494 [Abstract] [Full Text]  
• Huyer, G., Kistler, A., Nouvet, F. J., George, C. M., Boyle, M. L., Michaelis, S. (2006). Saccharomyces cerevisiae a-Factor Mutants Reveal Residues Critical for Processing, Activity, and Export.. Eukaryot Cell 5: 1560-1570 [Abstract] [Full Text]  
• Young, S. G., Fong, L. G., Michaelis, S. (2005). Thematic Review Series: Lipid Posttranslational Modifications. Prelamin A, Zmpste24, misshapen cell nuclei, and progeria--new evidence suggesting that protein farnesylation could be important for disease pathogenesis. J. Lipid Res. 46: 2531-2558 [Abstract] [Full Text]  
• Kim, S., Lapham, A. N., Freedman, C. G. K., Reed, T. L., Schmidt, W. K. (2005). Yeast as a Tractable Genetic System for Functional Studies of the Insulin-degrading Enzyme. J. Biol. Chem. 280: 27481-27490 [Abstract] [Full Text]  
• Riquelme, M., Challen, M. P., Casselton, L. A., Brown, A. J. (2005). The Origin of Multiple B Mating Specificities in Coprinus cinereus. Genetics 170: 1105-1119 [Abstract] [Full Text]  
• Yamashita, A., Kamata, R., Kawagishi, N., Nakanishi, H., Suzuki, H., Sugiura, T., Waku, K. (2005). Roles of C-Terminal Processing, and Involvement in Transacylation Reaction of Human Group IVC Phospholipase A2 (cPLA2{gamma}). J Biochem 137: 557-567 [Abstract] [Full Text]  
• Fabre, E., Muller, H., Therizols, P., Lafontaine, I., Dujon, B., Fairhead, C. (2005). Comparative Genomics in Hemiascomycete Yeasts: Evolution of Sex, Silencing, and Subtelomeres. Mol Biol Evol 22: 856-873 [Abstract] [Full Text]  
• Dresselhaus, T., Amien, S., Marton, M., Strecke, A., Brettschneider, R., Cordts, S. (2005). TRANSPARENT LEAF AREA1 Encodes a Secreted Proteolipid Required for Anther Maturation, Morphogenesis, and Differentiation during Leaf Development in Maize. Plant Cell 17: 730-745 [Abstract] [Full Text]  
• Hsueh, Y.-P., Shen, W.-C. (2005). A Homolog of Ste6, the a-Factor Transporter in Saccharomyces cerevisiae, Is Required for Mating but Not for Monokaryotic Fruiting in Cryptococcus neoformans. Eukaryot Cell 4: 147-155 [Abstract] [Full Text]  
• Huyer, G., Longsworth, G. L., Mason, D. L., Mallampalli, M. P., McCaffery, J. M., Wright, R. L., Michaelis, S. (2004). A Striking Quality Control Subcompartment in Saccharomyces cerevisiae: The Endoplasmic Reticulum-associated Compartment. Mol. Biol. Cell 15: 908-921 [Abstract] [Full Text]  
• Mason, D. L., Mallampalli, M. P., Huyer, G., Michaelis, S. (2003). A Region within a Lumenal Loop of Saccharomyces cerevisiae Ycf1p Directs Proteolytic Processing and Substrate Specificity. Eukaryot Cell 2: 588-598 [Abstract] [Full Text]  
• Mason, D. L., Michaelis, S. (2002). Requirement of the N-Terminal Extension for Vacuolar Trafficking and Transport Activity of Yeast Ycf1p, an ATP-binding Cassette Transporter. Mol. Biol. Cell 13: 4443-4455 [Abstract] [Full Text]  
• Shen, W.-C., Davidson, R. C., Cox, G. M., Heitman, J. (2002). Pheromones Stimulate Mating and Differentiation via Paracrine and Autocrine Signaling in Cryptococcus neoformans. Eukaryot Cell 1: 366-377 [Abstract] [Full Text]  
• Sharma, K. G., Mason, D. L., Liu, G., Rea, P. A., Bachhawat, A. K., Michaelis, S. (2002). Localization, Regulation, and Substrate Transport Properties of Bpt1p, a Saccharomyces cerevisiae MRP-Type ABC Transporter. Eukaryot Cell 1: 391-400 [Abstract] [Full Text]  
• McClelland, C. M., Fu, J., Woodlee, G. L., Seymour, T. S., Wickes, B. L. (2002). Isolation and Characterization of the Cryptococcus neoformans MATa Pheromone Gene. Genetics 160: 935-947 [Abstract] [Full Text]  
• Schneider, K. B., Palmer, T. M., Grossman, A. D. (2002). Characterization of comQ and comX, Two Genes Required for Production of ComX Pheromone in Bacillus subtilis. J. Bacteriol. 184: 410-419 [Abstract] [Full Text]  
• Tam, A., Schmidt, W. K., Michaelis, S. (2001). The Multispanning Membrane Protein Ste24p Catalyzes CAAX Proteolysis and NH2-terminal Processing of the Yeast a-Factor Precursor. J. Biol. Chem. 276: 46798-46806 [Abstract] [Full Text]  
• Schmidt, W. K., Tam, A., Michaelis, S. (2000). Reconstitution of the Ste24p-dependent N-terminal Proteolytic Step in Yeast a-Factor Biogenesis. J. Biol. Chem. 275: 6227-6233 [Abstract] [Full Text]  
• Bi, E., Chiavetta, J. B., Chen, H., Chen, G.-C., Chan, C. S. M., Pringle, J. R. (2000). Identification of Novel, Evolutionarily Conserved Cdc42p-interacting Proteins and of Redundant Pathways Linking Cdc24p and Cdc42p to Actin Polarization in Yeast. Mol. Biol. Cell 11: 773-793 [Abstract] [Full Text]  
• Meacham, G. C., Browne, B. L., Zhang, W., Kellermayer, R., Bedwell, D. M., Cyr, D. M. (1999). Mutations in the Yeast Hsp40 Chaperone Protein Ydj1 Cause Defects in Axl1 Biogenesis and Pro-a-factor Processing. J. Biol. Chem. 274: 34396-34402 [Abstract] [Full Text]  
• Gaudry, C. A., Verderio, E., Aeschlimann, D., Cox, A., Smith, C., Griffin, M. (1999). Cell Surface Localization of Tissue Transglutaminase Is Dependent on a Fibronectin-binding Site in Its N-terminal beta -Sandwich Domain. J. Biol. Chem. 274: 30707-30714 [Abstract] [Full Text]  
• Fowler, T. J., DeSimone, S. M., Mitton, M. F., Kurjan, J., Raper, C. A. (1999). Multiple Sex Pheromones and Receptors of a Mushroom-producing Fungus Elicit Mating in Yeast. Mol. Biol. Cell 10: 2559-2572 [Abstract] [Full Text]  
• Elia, L., Marsh, L. (1998). A Role for a Protease in Morphogenic Responses during Yeast Cell Fusion. JCB 142: 1473-1485 [Abstract] [Full Text]  
• Schmidt, W. K., Tam, A., Fujimura-Kamada, K., Michaelis, S. (1998). Endoplasmic reticulum membrane localization of Rce1p and Ste24p, yeast proteases involved in carboxyl-terminal CAAX protein processing and amino-terminal a-factor cleavage. Proc. Natl. Acad. Sci. USA 95: 11175-11180 [Abstract] [Full Text]  
• Boyartchuk, V. L., Rine, J. (1998). Roles of Prenyl Protein Proteases in Maturation of Saccharomyces cerevisiae a-Factor. Genetics 150: 95-101 [Abstract] [Full Text]  
• Tam, A., Nouvet, F. J., Fujimura-Kamada, K., Slunt, H., Sisodia, S. S., Michaelis, S. (1998). Dual Roles for Ste24p in Yeast a-Factor Maturation: NH2-terminal Proteolysis and COOH-terminal CAAX Processing. JCB 142: 635-649 [Abstract] [Full Text]  
• Romano, J. D., Schmidt, W. K., Michaelis, S. (1998). The Saccharomyces cerevisiae Prenylcysteine Carboxyl Methyltransferase Ste14p Is in the Endoplasmic Reticulum Membrane. Mol. Biol. Cell 9: 2231-2247 [Abstract] [Full Text]  
• Stanley, P., Koronakis, V., Hughes, C. (1998). Acylation of Escherichia coli Hemolysin: A Unique Protein Lipidation Mechanism Underlying Toxin Function. Microbiol. Mol. Biol. Rev. 62: 309-333 [Abstract] [Full Text]  
• Fujimura-Kamada, K., Nouvet, F. J., Michaelis, S. (1997). A Novel Membrane-associated Metalloprotease, Ste24p, Is Required for the First Step of NH2-terminal Processing of the Yeast a-Factor Precursor. JCB 136: 271-285 [Abstract] [Full Text]  
• Leung, G. K., Schmidt, W. K., Bergo, M. O., Gavino, B., Wong, D. H., Tam, A., Ashby, M. N., Michaelis, S., Young, S. G. (2001). Biochemical Studies of Zmpste24-deficient Mice. J. Biol. Chem. 276: 29051-29058 [Abstract] [Full Text]  

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