A genetically encoded Förster resonance energy transfer sensor for monitoring in vivo trehalose-6-phosphate dynamics |
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Authors: | Estevão A Peroza Jennifer C Ewald Geetha Parakkal Jan M Skotheim Nicola Zamboni |
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Institution: | 1. Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland;2. Department of Biology, Stanford University, Stanford, CA 94305, USA |
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Abstract: | Trehalose-6-phosphate is a pivotal regulator of sugar metabolism, growth, and osmotic equilibrium in bacteria, yeasts, and plants. To directly visualize the intracellular levels of intracellular trehalose-6-phosphate, we developed a series of specific Förster resonance energy transfer (FRET) sensors for in vivo microscopy. We demonstrated real-time monitoring of regulation in the trehalose pathway of Escherichia coli. In Saccharomyces cerevisiae, we could show that the concentration of free trehalose-6-phosphate during growth on glucose is in a range sufficient for inhibition of hexokinase. These findings support the hypothesis of trehalose-6-phosphate as the effector of a negative feedback system, similar to the inhibition of hexokinase by glucose-6-phosphate in mammalian cells and controlling glycolytic flux. |
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Keywords: | Trehalose-6-phosphate FRET sensor Glycolysis regulation |
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