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A yeast FRET biosensor enlightens cAMP signaling.
- Source :
-
Molecular biology of the cell [Mol Biol Cell] 2021 Jun 15; Vol. 32 (13), pp. 1229-1240. Date of Electronic Publication: 2021 Apr 21. - Publication Year :
- 2021
-
Abstract
- The cAMP-PKA signaling cascade in budding yeast regulates adaptation to changing environments. We developed yEPAC, a FRET-based biosensor for cAMP measurements in yeast. We used this sensor with flow cytometry for high-throughput single cell-level quantification during dynamic changes in response to sudden nutrient transitions. We found that the characteristic cAMP peak differentiates between different carbon source transitions and is rather homogenous among single cells, especially for transitions to glucose. The peaks are mediated by a combination of extracellular sensing and intracellular metabolism. Moreover, the cAMP peak follows the Weber-Fechner law; its height scales with the relative, and not the absolute, change in glucose. Last, our results suggest that the cAMP peak height conveys information about prospective growth rates. In conclusion, our yEPAC-sensor makes possible new avenues for understanding yeast physiology, signaling, and metabolic adaptation.
- Subjects :
- Biosensing Techniques methods
Cyclic AMP metabolism
Cyclic AMP-Dependent Protein Kinases metabolism
Flow Cytometry methods
Glucose metabolism
High-Throughput Screening Assays methods
Prospective Studies
Saccharomyces cerevisiae metabolism
Saccharomyces cerevisiae Proteins metabolism
Signal Transduction physiology
Single-Cell Analysis methods
Cyclic AMP analysis
Cyclic AMP-Dependent Protein Kinases analysis
Fluorescence Resonance Energy Transfer methods
Subjects
Details
- Language :
- English
- ISSN :
- 1939-4586
- Volume :
- 32
- Issue :
- 13
- Database :
- MEDLINE
- Journal :
- Molecular biology of the cell
- Publication Type :
- Academic Journal
- Accession number :
- 33881352
- Full Text :
- https://doi.org/10.1091/mbc.E20-05-0319