AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions

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. As a proof of principle, we show in both cell lines and primary cell cultu...

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Bibliographic Details
Published in:	BMC biology 2020-07, Vol.18 (1), p.1-81, Article 81
Main Authors:	Bouquier, Nathalie, Moutin, Enora, Tintignac, Lionel A, Reverbel, Amandine, Jublanc, Elodie, Sinnreich, Michael, Chastagnier, Yan, Averous, Julien, Fafournoux, Pierre, Verpelli, Chiara, Boeckers, Tobias, Carnac, Gilles, Perroy, Julie, Ollendorff, Vincent
Format:	Article
Language:	eng
Subjects:	Amino acids Animal models Antibiotics Autism Autophagy Biochemistry, Molecular Biology Biopsy Biosensors Biosynthesis Biotechnology BRET Cell differentiation Cell growth Cell lines Cells (biology) Cytosol Deoxyribonucleic acid Detection equipment Detectors Differentiation (biology) DNA Energy transfer Genetic code Growth factors Hippocampus Insulin Kinases Leucine Life Sciences Metabolism Mitochondria mTor signaling mTORC1 Biosensor mToropathies Muscle differentiation Muscles Neuronal activity Neurons Peptides Phenotypes Phosphorylation Plasmids Proteins Signaling TOR protein
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Summary:	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. 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. Altogether, our results demonstrate that AIMTOR is a sensitive and specific tool to investigate mTOR-signaling dynamics in living cells and phenotype mTORopathies.
ISSN:	1741-7007 1741-7007