, 2008; Nakase et al., 2008). FSM development requires the secretory pathway and vesicle docking (Shimoda, 2004; Shimoda & Nakamura, 2004; Nakamura et al., 2008). Spores maturate with the building of a specialized cell STA-9090 supplier wall, which involves the synthesis of α- and β-glucan and chitin (Arellano et al., 2000; Liu et al., 2000; Martín et al., 2000; Matsuo et al., 2005; Garcia et al., 2006; de Medina-Redondo

et al., 2008). The exocyst is a protein complex involved in the tethering and spatial targeting of post-Golgi vesicles to the plasma membrane before vesicle fusion (TerBush et al., 1996; Guo et al., 1999; Mehta et al., 2005). In S. pombe, this complex participates in cell separation because it is required to target hydrolytic enzymes to the septum (Wang et al., 2002; Martin-Cuadrado

et al., 2005). The only viable exocyst mutants in this organism are sec8-1 and exo70Δ (Wang et al., 2002, 2003). The term exomer refers to a Saccharomyces cerevisiae coat complex required for the transport of certain membrane proteins from the trans-Golgi network to the plasma membrane (Wang et al., 2006; Barfield et al., 3-MA in vivo 2009). The exomer subunit Chs5p is required for chitin synthesis and mating (Santos et al., 1997). In S. pombe, the Chs5p-homologue Cfr1p is required for cell wall digestion during mating (Cartagena-Lirola et al., 2006). The analysis of how cell wall-modifying enzymes required for sexual development reach the cell surface is not only interesting for the characterization of the mating process in yeast but also represents a model system to study intracellular trafficking during a developmental process. The initial goal of this work was to study the regulation of cell adhesion by genes that have already been implicated in the mating and/or the cell wall remodeling

processes. To do so, we analyzed agglutination in several mutants; the mutants selected were spk1Δ (defective in the mating signal transduction pathway; Nielsen, 2004), spm1Δ (deleted for a MAP kinase that regulates morphogenesis, cell integrity, and mating; Zaitsevskaya-Carter & Cooper, 1997), dni1Δ (deleted for a claudin-like tetraspan protein required for cell wall reorganization and membrane fusion during mating; Clemente-Ramos et al., 2009), cfr1Δ (deleted for an exomer component; Cartagena-Lirola et al., 2006), sec8-1 (bearing Astemizole a point mutation in sec8+; Wang et al., 2002), and exo70Δ (deleted for exo70+; Wang et al., 2003). Surprisingly, our results showed that agglutination is dependent on Sec8p, but independent of Exo70p. This result prompted us to analyze in detail the role of these exocyst subunits in mating. Our results suggest that Sec8p and Exo70p participate in different subcomplexes that are differentially required during sexual development. All techniques for S. pombe growth and manipulation have been described elsewhere (http://www.biotwiki.org/bin/view/Pombe/NurseLabManual). The relevant genotype of the strains used is listed in Supporting Information, Table S1.