AIMTOR, a BRET biosensor for live imaging, reveals subcellular mTOR signaling and dysfunctions
Autor: | Lionel Tintignac, Elodie Jublanc, Enora Moutin, Nathalie Bouquier, Julien Averous, Yan Chastagnier, Michael Sinnreich, Amandine Reverbel, Chiara Verpelli, Vincent Ollendorff, Gilles Carnac, Tobias M. Boeckers, Julie Perroy, Pierre Fafournoux |
---|---|
Přispěvatelé: | Institut de Génomique Fonctionnelle (IGF), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre National de la Recherche Scientifique (CNRS), University Hospital Basel [Basel], Dynamique Musculaire et Métabolisme (DMEM), Université de Montpellier (UM)-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Unité de Nutrition Humaine (UNH), Université Clermont Auvergne [2017-2020] (UCA [2017-2020])-Institut National de Recherche pour l’Agriculture, l’Alimentation et l’Environnement (INRAE), Institute of Neurosciences, AMRI Hospitals, Universität Ulm - Ulm University [Ulm, Allemagne], Physiologie & médecine expérimentale du Cœur et des Muscles [U 1046] (PhyMedExp), European Research Council (ERC) 646788, grant of INRAE (department ALIMH), grant of INRAE (department PHASE), Region OccitanieRegion Ile-de-France, ANR-13-JSV4-0005,SYN CITY,Remodelage des complexes d'échafaudage: Dynamique et fonction du réceptosome de mGlu5 dans la plasticité synaptique(2013), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), MORNET, Dominique, Jeunes Chercheuses et Jeunes Chercheurs - Remodelage des complexes d'échafaudage: Dynamique et fonction du réceptosome de mGlu5 dans la plasticité synaptique - - SYN CITY2013 - ANR-13-JSV4-0005 - JC - VALID |
Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Diagnostic Imaging
[SDV.BIO]Life Sciences [q-bio]/Biotechnology Physiology mTor signaling Biosensing Techniques Plant Science mTORC1 Mitochondrion Biology General Biochemistry Genetics and Molecular Biology Quadriceps Muscle Mice 03 medical and health sciences 0302 clinical medicine Structural Biology Live cell imaging Lysosome [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology medicine Animals Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Autism spectrum disorder lcsh:QH301-705.5 Ecology Evolution Behavior and Systematics PI3K/AKT/mTOR pathway 030304 developmental biology Muscle differentiation 0303 health sciences Neuronal activity mTORC1 Biosensor Muscle cell differentiation TOR Serine-Threonine Kinases Cell Biology [SDV.BIO] Life Sciences [q-bio]/Biotechnology Cell biology Cytosol HEK293 Cells medicine.anatomical_structure lcsh:Biology (General) Phosphorylation BRET General Agricultural and Biological Sciences 030217 neurology & neurosurgery mToropathies Signal Transduction Research Article Developmental Biology Biotechnology |
Zdroj: | BMC Biology, Vol 18, Iss 1, Pp 1-17 (2020) BMC Biology BMC Biology, 2020, 18 (1), pp.81. ⟨10.1186/s12915-020-00790-8⟩ BMC Biology, BioMed Central, 2020, 18 (1), pp.81. ⟨10.1186/s12915-020-00790-8⟩ |
ISSN: | 1741-7007 |
Popis: | BackgroundmTOR 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.ResultsAs 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.ConclusionsAltogether, our results demonstrate that AIMTOR is a sensitive and specific tool to investigate mTOR-signaling dynamics in living cells and phenotype mTORopathies. |
Databáze: | OpenAIRE |
Externí odkaz: | |
Nepřihlášeným uživatelům se plný text nezobrazuje | K zobrazení výsledku je třeba se přihlásit. |