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Temporal and Spatial Properties of a Yeast Multi-Cellular Amplification System Based on Signal Molecule Diffusion
Institute of Genetics, Technische Universität Dresden, Helmholtzstr. 10, 01062 Dresden, Germany
Helmholtz Centre for Environmental Research UFZ, Department for Environmental Microbiology, Permoserstr. 15, 04318 Leipzig, Germany
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 22 August 2013; in revised form: 11 October 2013 / Accepted: 18 October 2013 / Published: 25 October 2013
Abstract: We report on the spatial and temporal signaling properties of a yeast pheromone-based cell communication and amplifier system. It utilizes the Saccharomyces cerevisiae mating response pathway and relies on diffusion of the pheromone α–factor as key signaling molecule between two cell types. One cell type represents the α–factor secreting sensor part and the other the reporter part emitting fluorescence upon activation. Although multi-cellular signaling systems promise higher specificity and modularity, the complex interaction of the cells makes prediction of sensor performance difficult. To test the maximum distance and response time between sensor and reporter cells, the two cell types were spatially separated in defined compartments of agarose hydrogel (5 ´ 5 mm) and reconnected by diffusion of the yeast pheromone. Different ratios of sensor to reporter cells were tested to evaluate the minimum amount of sensor cells required for signal transduction. Even the smallest ratio, one α–factor-secreting cell to twenty reporter cells, generated a distinct fluorescence signal. When using a 1:1 ratio, the secreted pheromone induced fluorescence in a distance of up to four millimeters after six hours. We conclude from both our experimental results and a mathematical diffusion model that in our approach: (1) the maximum dimension of separated compartments should not exceed five millimeters in gradient direction; and (2) the time-limiting step is not diffusion of the signaling molecule but production of the reporter protein.
Keywords: microbial biosensor; yeast; alpha (α)–factor; fluorescence; immobilization; agarose; modular signaling/amplification system
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Jahn, M.; Mölle, A.; Rödel, G.; Ostermann, K. Temporal and Spatial Properties of a Yeast Multi-Cellular Amplification System Based on Signal Molecule Diffusion. Sensors 2013, 13, 14511-14522.
Jahn M, Mölle A, Rödel G, Ostermann K. Temporal and Spatial Properties of a Yeast Multi-Cellular Amplification System Based on Signal Molecule Diffusion. Sensors. 2013; 13(11):14511-14522.
Jahn, Michael; Mölle, Annett; Rödel, Gerhard; Ostermann, Kai. 2013. "Temporal and Spatial Properties of a Yeast Multi-Cellular Amplification System Based on Signal Molecule Diffusion." Sensors 13, no. 11: 14511-14522.