TY - JOUR
T1 - A yeast FRET biosensor enlightens cAMP signaling
AU - Botman, Dennis
AU - O'Toole, Tom G.
AU - Goedhart, Joachim
AU - Bruggeman, Frank J.
AU - van Heerden, Johan H.
AU - Teusink, Bas
N1 - Funding Information:
We thank Kees Jalink (Division of Cell Biology, The Netherlands Cancer Institute) for sharing mTurquoise2Δ-Epac(CD,ΔDEP)-cp173Venus-cp173Venus (Epac-SH188). We thank Juan Garcia Vallejo and Cora Chadick (Molecular Cell Biology & Immunology, VUmc) for his help with flow cytometry. We are also grateful to Sonia Colombo and V.O. Nikolaev for sharing YFP-EPAC2-CFP with us. We thank Joris Winderickx (Functional Biology, KU Leuven) and Marco Siderius (Amsterdam Institute for Molecules, Medicines and Systems [AIMMS], Division of Medicinal Chemistry) for sharing W303-1A mutant strains. We thank Daan de Groot and Philipp Sa-vakis for fruitful discussions. This work was supported by the NWO VICI grant 865.14.005
Publisher Copyright:
© 2021 Botman et al.
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
PY - 2021/6/15
Y1 - 2021/6/15
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85108386575&partnerID=8YFLogxK
U2 - 10.1091/MBC.E20-05-0319
DO - 10.1091/MBC.E20-05-0319
M3 - Article
C2 - 33881352
VL - 32
SP - 1229
EP - 1240
JO - Molecular Biology of the Cell
JF - Molecular Biology of the Cell
SN - 1059-1524
IS - 13
ER -