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Article Dans Une Revue BMC Biology Année : 2020

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

Enora Moutin
Elodie Jublanc
Julien Averous
Pierre Fafournoux
Julie Perroy

Résumé

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.
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hal-02889781 , version 1 (22-07-2020)

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Nathalie Bouquier, Enora Moutin, Lionel J. Tintignac, Amandine Reverbel, Elodie Jublanc, et al.. AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions. BMC Biology, 2020, 18 (1), pp.81. ⟨10.1186/s12915-020-00790-8⟩. ⟨hal-02889781⟩
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