Dominant negative alleles of SEC10 reveal distinct domains involved in secretion and morphogenesis in yeast

• Published in: Molecular biology of the cell Vol. 9; no. 7; pp. 1725 - 1739
• Main Authors: Roth, D, Guo, W, Novick, P
• Format: Journal Article
• Language: English
• Published: United States The American Society for Cell Biology 01.07.1998
• Subjects: Alleles; Fungal Proteins - biosynthesis; Fungal Proteins - genetics; Fungal Proteins - metabolism; Genes, Dominant; Genes, Fungal; Genes, Lethal - genetics; GTP-Binding Proteins - biosynthesis; GTP-Binding Proteins - metabolism; Mutagenesis, Site-Directed; Myosin Heavy Chains; Myosin Type V; Myosins - metabolism; Organelles - physiology; Peptide Fragments - biosynthesis; Peptide Fragments - genetics; Peptide Fragments - metabolism; Phenotype; rab GTP-Binding Proteins; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - growth & development; Saccharomyces cerevisiae - physiology; Saccharomyces cerevisiae Proteins; Vesicular Transport Proteins
• ISSN: 1059-1524; 1939-4586
• DOI: 10.1091/mbc.9.7.1725

The accurate targeting of secretory vesicles to distinct sites on the plasma membrane is necessary to achieve polarized growth and to establish specialized domains at the surface of eukaryotic cells. Members of a protein complex required for exocytosis, the exocyst, have been localized to regions of active secretion in the budding yeast Saccharomyces cerevisiae where they may function to specify sites on the plasma membrane for vesicle docking and fusion. In this study we have addressed the function of one member of the exocyst complex, Sec10p. We have identified two functional domains of Sec10p that act in a dominant-negative manner to inhibit cell growth upon overexpression. Phenotypic and biochemical analysis of the dominant-negative mutants points to a bifunctional role for Sec10p. One domain, consisting of the amino-terminal two-thirds of Sec10p directly interacts with Sec15p, another exocyst component. Overexpression of this domain displaces the full-length Sec10 from the exocyst complex, resulting in a block in exocytosis and an accumulation of secretory vesicles. The carboxy-terminal domain of Sec10p does not interact with other members of the exocyst complex and expression of this domain does not cause a secretory defect. Rather, this mutant results in the formation of elongated cells, suggesting that the second domain of Sec10p is required for morphogenesis, perhaps regulating the reorientation of the secretory pathway from the tip of the emerging daughter cell toward the mother-daughter connection during cell cycle progression.