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Free keywords:
yeast; signal; mating factor; ELISA
Abstract:
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 signalling cascade that
has become a model for studying signal transduction. In contrast, the factors
contributing to the production of pheromone itself are poorly characterized,
despite the widespread use of the S. cerevisiae a-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 a-pheromone secretion. In the absence of pheromone from
the opposite mating type, we found that each cell secretes over 550 mature apheromone
peptides per second; 90% of this total was produced from MFa1.
The addition of a-pheromone more than doubled total a-pheromone secretion.
This technique offers several improvements on current methods for measuring
a-pheromone production and will allow detailed investigation of the factors
regulating pheromone production in yeast.
