Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA
A major signal transduction pathway regulating cell growth and many associated physiological properties as a function of nutrient availability in the yeast Saccharomyces cerevisiae is the protein kinase A (PKA) pathway. Glucose activation of PKA is mediated by G-protein coupled receptor (GPCR) Gpr1,...
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| Natura: | Articolo |
| Lingua: | English |
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Shared Science Publishers OG
2020-10-01
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| Serie: | Microbial Cell |
| Soggetti: | |
| Accesso online: | http://microbialcell.com/researcharticles/2020a-van-zeebroeck-microbial-cell/ |
| _version_ | 1826532060921069568 |
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| author | Griet Van Zeebroeck Liesbeth Demuyser Zhiqiang Zhang Ines Cottignie Johan M. Thevelein |
| author_facet | Griet Van Zeebroeck Liesbeth Demuyser Zhiqiang Zhang Ines Cottignie Johan M. Thevelein |
| author_sort | Griet Van Zeebroeck |
| collection | DOAJ |
| description | A major signal transduction pathway regulating cell growth and many associated physiological properties as a function of nutrient availability in the yeast Saccharomyces cerevisiae is the protein kinase A (PKA) pathway. Glucose activation of PKA is mediated by G-protein coupled receptor (GPCR) Gpr1, and secondary messenger cAMP. Other nutrients, including nitrogen, phosphate and sulfate, activate PKA in accordingly-starved cells through nutrient transceptors, but apparently without cAMP signaling. We have now used an optimized EPAC-based fluorescence resonance energy transfer (FRET) sensor to precisely monitor in vivo cAMP levels after nutrient addition. We show that GPCR-mediated glucose activation of PKA is correlated with a rapid transient increase in the cAMP level in vivo, whereas nutrient transceptor-mediated activation by nitrogen, phosphate or sulfate, is not associated with any significant increase in cAMP in vivo. We also demonstrate direct physical interaction between the Gap1 amino acid transceptor and the catalytic subunits of PKA, Tpk1, 2 and 3. In addition, we reveal a conserved consensus motif in the nutrient transceptors that is also present in Bcy1, the regulatory subunit of PKA. This suggests that nutrient transceptor activation of PKA may be mediated by direct release of bound PKA catalytic subunits, triggered by the conformational changes occurring during transport of the substrate by the transceptor. Our results support a model in which nutrient transceptors are evolutionary ancestors of GPCRs, employing a more primitive direct signaling mechanism compared to the indirect cAMP second-messenger signaling mechanism used by GPCRs for activation of PKA. |
| first_indexed | 2024-12-17T15:01:54Z |
| format | Article |
| id | doaj.art-c572f020463048d5896ba3b51e1b9e87 |
| institution | Directory Open Access Journal |
| issn | 2311-2638 |
| language | English |
| last_indexed | 2025-03-14T01:45:14Z |
| publishDate | 2020-10-01 |
| publisher | Shared Science Publishers OG |
| record_format | Article |
| series | Microbial Cell |
| spelling | doaj.art-c572f020463048d5896ba3b51e1b9e872025-03-12T13:46:43ZengShared Science Publishers OGMicrobial Cell2311-26382020-10-0181172710.15698/mic2021.01.740Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKAGriet Van Zeebroeck0Liesbeth Demuyser1Zhiqiang Zhang2Ines Cottignie3Johan M. Thevelein4Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, B-3001 Leuven-Heverlee, Flanders, Belgium.Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, B-3001 Leuven-Heverlee, Flanders, Belgium.Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, B-3001 Leuven-Heverlee, Flanders, Belgium.Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, B-3001 Leuven-Heverlee, Flanders, Belgium.Laboratory of Molecular Cell Biology, Institute of Botany and Microbiology, KU Leuven, B-3001 Leuven-Heverlee, Flanders, Belgium.A major signal transduction pathway regulating cell growth and many associated physiological properties as a function of nutrient availability in the yeast Saccharomyces cerevisiae is the protein kinase A (PKA) pathway. Glucose activation of PKA is mediated by G-protein coupled receptor (GPCR) Gpr1, and secondary messenger cAMP. Other nutrients, including nitrogen, phosphate and sulfate, activate PKA in accordingly-starved cells through nutrient transceptors, but apparently without cAMP signaling. We have now used an optimized EPAC-based fluorescence resonance energy transfer (FRET) sensor to precisely monitor in vivo cAMP levels after nutrient addition. We show that GPCR-mediated glucose activation of PKA is correlated with a rapid transient increase in the cAMP level in vivo, whereas nutrient transceptor-mediated activation by nitrogen, phosphate or sulfate, is not associated with any significant increase in cAMP in vivo. We also demonstrate direct physical interaction between the Gap1 amino acid transceptor and the catalytic subunits of PKA, Tpk1, 2 and 3. In addition, we reveal a conserved consensus motif in the nutrient transceptors that is also present in Bcy1, the regulatory subunit of PKA. This suggests that nutrient transceptor activation of PKA may be mediated by direct release of bound PKA catalytic subunits, triggered by the conformational changes occurring during transport of the substrate by the transceptor. Our results support a model in which nutrient transceptors are evolutionary ancestors of GPCRs, employing a more primitive direct signaling mechanism compared to the indirect cAMP second-messenger signaling mechanism used by GPCRs for activation of PKA.http://microbialcell.com/researcharticles/2020a-van-zeebroeck-microbial-cell/fret biosensorcamppkanutrient sensinggpcrtransceptoryeast |
| spellingShingle | Griet Van Zeebroeck Liesbeth Demuyser Zhiqiang Zhang Ines Cottignie Johan M. Thevelein Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA Microbial Cell fret biosensor camp pka nutrient sensing gpcr transceptor yeast |
| title | Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA |
| title_full | Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA |
| title_fullStr | Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA |
| title_full_unstemmed | Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA |
| title_short | Nutrient sensing and cAMP signaling in yeast: G-protein coupled receptor versus transceptor activation of PKA |
| title_sort | nutrient sensing and camp signaling in yeast g protein coupled receptor versus transceptor activation of pka |
| topic | fret biosensor camp pka nutrient sensing gpcr transceptor yeast |
| url | http://microbialcell.com/researcharticles/2020a-van-zeebroeck-microbial-cell/ |
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