Application of the yeast pheromone system for controlled cell–cell communication and signal amplification

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Aims:The aim of the work is to exploit the yeast pheromone system for controlled cell–cell communication and as an amplification circuit in technical applications, e.g. biosensors or sensor-actor systems.Methods and Results:As a proof of principle, we developed recombinant Saccharomyces cerevisiae cells that express enhanced green fluorescent protein (EGFP) in response to different concentrations of the alpha (α)-factor mating pheromone. A respective reporter construct allowing the pheromone-driven expression of EGFP was transformed into the S. cerevisiae strains BY4741 and BY4741 bar1Δ. Upon addition of synthetic α-factor, the fluorescence strongly increases after 4 h. Furthermore, cells with constitutive α-factor expression were able to induce the expression of EGFP in co-cultivation with sensor cells only if both cell types were deleted for the gene BAR1, encoding α-factor protease. For technical applications, the immobilization of functionalized cells may be beneficial. We show that pheromone-induced expression of EGFP is effective in alginate-immobilized cells.Conclusions:Based on S. cerevisiaeα-factor, we developed a controlled cell–cell communication system and amplification circuit for pheromone-driven expression of a target protein. The system is effective both in suspension and after cell immobilization.Significance and Impact of the Study:The developed set of recombinant yeast strains is the basis to apply the yeast pheromone system for signal production and amplification in biosensors or sensor-actor systems.

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