title: A genetically encoded Förster resonance energy transfer sensor for monitoring in vivo trehalose-6-phosphate dynamics.
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.
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Estevão A
authors.initials: EA
authors.lastname: Peroza
authors.affiliation: Department of Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Ewald
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Geetha
authors.initials: G
authors.lastname: Parakkal
authors.affiliation: Department of Biology, Stanford University, Stanford, CA 94305, USA.
authors.forename: Jan M
authors.initials: JM
authors.lastname: Skotheim
authors.affiliation: Institute of Molecular Systems Biology, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Nicola
authors.initials: N
authors.lastname: Zamboni
mesh_terms: D015374:Biosensing Techniques; D004926:Escherichia coli / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005821:Genetic Techniques; D008164:Luminescent Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D013403:Sugar Phosphates / Q000378:metabolism*; D014199:Trehalose / Q000031:analogs & derivatives* / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D013403:Sugar Phosphates; C082722:trehalose-6-phosphate; D014199:Trehalose
keywords: FRET sensor; Glycolysis regulation; Trehalose-6-phosphate