Molecular quantification of Saccharomyces cerevisiae α-pheromone secretion.

SUMMARY: Saccharomyces cerevisiae yeast cells court each other by producing an attractive sex pheromone specific to their mating-type. Cells detect the sex pheromone from potential mates using a well-defined intracellular signaling cascade that has become a model for studying signal transduction. In contrast, the factors contributing to production of pheromone ...
itself are poorly characterized, despite the widespread use of the S. cerevisiae α-pheromone secretion pathway in industrial fungal protein expression systems. Progress in understanding pheromone secretion has been hindered by a lack of a precise and quantitative pheromone production assay. Here, we present an ELISA-based method for the quantification of α-pheromone secretion. In the absence of pheromone from the opposite mating type, we found that each cell secretes over 550 mature α-pheromone peptides per second; 90% of this total was produced from MFα1. The addition of a-pheromone more than doubled total α-pheromone secretion. This technique offers several improvements on current methods for measuring α-pheromone production, and will allow detailed investigation of the factors regulating pheromone production in yeast. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
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Date: Jun. 04, 2012
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