Localized deposition of chitin on the yeast cell surface in response to mating pheromone.
Treatment of a mating-type Saccharomyces cerevisiae cells with the pheromone alpha-factor (secreted by alpha mating-type cells) induces the synthesis of chitin. Small daughter cells, which start with no detectable chitin, make 3 times more chitin when grown in the presence of alpha-factor than do untreated exponentially growing cells. Budding cells accumulate chitin in the nascent division septum [Cabib, E. & Bowers, B. (1975) J. Bacteriol. 124, 1586), as detected by staining with the fluorescent dye primulin. In the absence of a division septum, alpha-factor-treated cells accumulate chitin in the area of pheromone-stimulated growth. Enzymatic lysis of budding and pheromone-treated cells allows the separation of membrane-bound chitin synthase (UDP-2-acetamido-2-deoxy-D-glucose: chitin 4-beta-acetamidodeoxyglucosyltransferase, EC 2.4.1.16) activity from a dense particulate fraction containing chitin. Chitin synthase activity is associated with both the plasma membrane and small intracellular particles. During pheromone treatment, the levels of chitin synthase in the plasma membrane and in intracellular particle fractions increase 11- and 4-fold, respectively. Although chitin synthase is made as zymogen that requires proteolytic activation, the plasma membrane of pheromone-treated cells shows a significant fraction of preactivated enzyme; intracellular membrane-bound synthase is found exclusively in the zymogen form.
Schekman
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Department of Biochemistry, University of California, Berkeley, California 94720.
Brawley
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