AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions
Abstract Background mTOR signaling is an essential nutrient and energetic sensing pathway. Here we describe AIMTOR, a sensitive genetically encoded BRET (Bioluminescent Resonance Energy Transfer) biosensor to study mTOR activity in living cells. Results As a proof of principle, we show in both cell...
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doaj-de858ec3955d4d94a438dc5be235f72e2020-11-25T02:50:13ZengBMCBMC Biology1741-70072020-07-0118111710.1186/s12915-020-00790-8AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctionsNathalie Bouquier0Enora Moutin1Lionel A. Tintignac2Amandine Reverbel3Elodie Jublanc4Michael Sinnreich5Yan Chastagnier6Julien Averous7Pierre Fafournoux8Chiara Verpelli9Tobias Boeckers10Gilles Carnac11Julie Perroy12Vincent Ollendorff13IGF, University of Montpellier, CNRS, INSERMIGF, University of Montpellier, CNRS, INSERMUniversity Hospital Basel, Department of BiomedecineDMEM, University of Montpellier, INRAEDMEM, University of Montpellier, INRAEUniversity Hospital Basel, Department of BiomedecineIGF, University of Montpellier, CNRS, INSERMUniversité Clermont Auvergne, INRAE, Unité de Nutrition Humaine, UMR1019Université Clermont Auvergne, INRAE, Unité de Nutrition Humaine, UMR1019Cnr Institute of NeuroscienceAnatomie und Zellbiologie Universität UlmPhymedexp, University of Montpellier, CNRS, INSERMIGF, University of Montpellier, CNRS, INSERMDMEM, University of Montpellier, INRAEAbstract Background mTOR signaling is an essential nutrient and energetic sensing pathway. Here we describe AIMTOR, a sensitive genetically encoded BRET (Bioluminescent Resonance Energy Transfer) biosensor to study mTOR activity in living cells. Results As a proof of principle, we show in both cell lines and primary cell cultures that AIMTOR BRET intensities are modified by mTOR activity changes induced by specific inhibitors and activators of mTORC1 including amino acids and insulin. We further engineered several versions of AIMTOR enabling subcellular-specific assessment of mTOR activities. We then used AIMTOR to decipher mTOR signaling in physio-pathological conditions. First, we show that mTORC1 activity increases during muscle cell differentiation and in response to leucine stimulation in different subcellular compartments such as the cytosol and at the surface of the lysosome, the nucleus, and near the mitochondria. Second, in hippocampal neurons, we found that the enhancement of neuronal activity increases mTOR signaling. AIMTOR further reveals mTOR-signaling dysfunctions in neurons from mouse models of autism spectrum disorder. Conclusions Altogether, our results demonstrate that AIMTOR is a sensitive and specific tool to investigate mTOR-signaling dynamics in living cells and phenotype mTORopathies.http://link.springer.com/article/10.1186/s12915-020-00790-8mTor signalingmTORC1 BiosensorBRETMuscle differentiationmToropathiesNeuronal activity |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Nathalie Bouquier Enora Moutin Lionel A. Tintignac Amandine Reverbel Elodie Jublanc Michael Sinnreich Yan Chastagnier Julien Averous Pierre Fafournoux Chiara Verpelli Tobias Boeckers Gilles Carnac Julie Perroy Vincent Ollendorff |
spellingShingle |
Nathalie Bouquier Enora Moutin Lionel A. Tintignac Amandine Reverbel Elodie Jublanc Michael Sinnreich Yan Chastagnier Julien Averous Pierre Fafournoux Chiara Verpelli Tobias Boeckers Gilles Carnac Julie Perroy Vincent Ollendorff AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions BMC Biology mTor signaling mTORC1 Biosensor BRET Muscle differentiation mToropathies Neuronal activity |
author_facet |
Nathalie Bouquier Enora Moutin Lionel A. Tintignac Amandine Reverbel Elodie Jublanc Michael Sinnreich Yan Chastagnier Julien Averous Pierre Fafournoux Chiara Verpelli Tobias Boeckers Gilles Carnac Julie Perroy Vincent Ollendorff |
author_sort |
Nathalie Bouquier |
title |
AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions |
title_short |
AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions |
title_full |
AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions |
title_fullStr |
AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions |
title_full_unstemmed |
AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions |
title_sort |
aimtor, a bret biosensor for live imaging, reveals subcellular mtor signaling and dysfunctions |
publisher |
BMC |
series |
BMC Biology |
issn |
1741-7007 |
publishDate |
2020-07-01 |
description |
Abstract Background mTOR signaling is an essential nutrient and energetic sensing pathway. Here we describe AIMTOR, a sensitive genetically encoded BRET (Bioluminescent Resonance Energy Transfer) biosensor to study mTOR activity in living cells. Results As a proof of principle, we show in both cell lines and primary cell cultures that AIMTOR BRET intensities are modified by mTOR activity changes induced by specific inhibitors and activators of mTORC1 including amino acids and insulin. We further engineered several versions of AIMTOR enabling subcellular-specific assessment of mTOR activities. We then used AIMTOR to decipher mTOR signaling in physio-pathological conditions. First, we show that mTORC1 activity increases during muscle cell differentiation and in response to leucine stimulation in different subcellular compartments such as the cytosol and at the surface of the lysosome, the nucleus, and near the mitochondria. Second, in hippocampal neurons, we found that the enhancement of neuronal activity increases mTOR signaling. AIMTOR further reveals mTOR-signaling dysfunctions in neurons from mouse models of autism spectrum disorder. Conclusions Altogether, our results demonstrate that AIMTOR is a sensitive and specific tool to investigate mTOR-signaling dynamics in living cells and phenotype mTORopathies. |
topic |
mTor signaling mTORC1 Biosensor BRET Muscle differentiation mToropathies Neuronal activity |
url |
http://link.springer.com/article/10.1186/s12915-020-00790-8 |
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