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Dynamics of mTORC1 activation in response to amino acids

Amino acids are essential activators of mTORC1 via a complex containing RAG GTPases, RAGULATOR and the vacuolar ATPase. Sensing of amino acids causes translocation of mTORC1 to lysosomes, an obligate step for activation. To examine the spatial and temporal dynamics of this translocation, we used liv...

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Autores principales: Manifava, Maria, Smith, Matthew, Rotondo, Sergio, Walker, Simon, Niewczas, Izabella, Zoncu, Roberto, Clark, Jonathan, Ktistakis, Nicholas T
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5059141/
https://www.ncbi.nlm.nih.gov/pubmed/27725083
http://dx.doi.org/10.7554/eLife.19960
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author Manifava, Maria
Smith, Matthew
Rotondo, Sergio
Walker, Simon
Niewczas, Izabella
Zoncu, Roberto
Clark, Jonathan
Ktistakis, Nicholas T
author_facet Manifava, Maria
Smith, Matthew
Rotondo, Sergio
Walker, Simon
Niewczas, Izabella
Zoncu, Roberto
Clark, Jonathan
Ktistakis, Nicholas T
author_sort Manifava, Maria
collection PubMed
description Amino acids are essential activators of mTORC1 via a complex containing RAG GTPases, RAGULATOR and the vacuolar ATPase. Sensing of amino acids causes translocation of mTORC1 to lysosomes, an obligate step for activation. To examine the spatial and temporal dynamics of this translocation, we used live imaging of the mTORC1 component RAPTOR and a cell permeant fluorescent analogue of di-leucine methyl ester. Translocation to lysosomes is a transient event, occurring within 2 min of aa addition and peaking within 5 min. It is temporally coupled with fluorescent leucine appearance in lysosomes and is sustained in comparison to aa stimulation. Sestrin2 and the vacuolar ATPase are negative and positive regulators of mTORC1 activity in our experimental system. Of note, phosphorylation of canonical mTORC1 targets is delayed compared to lysosomal translocation suggesting a dynamic and transient passage of mTORC1 from the lysosomal surface before targetting its substrates elsewhere. DOI: http://dx.doi.org/10.7554/eLife.19960.001
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spelling pubmed-50591412016-10-12 Dynamics of mTORC1 activation in response to amino acids Manifava, Maria Smith, Matthew Rotondo, Sergio Walker, Simon Niewczas, Izabella Zoncu, Roberto Clark, Jonathan Ktistakis, Nicholas T eLife Biochemistry Amino acids are essential activators of mTORC1 via a complex containing RAG GTPases, RAGULATOR and the vacuolar ATPase. Sensing of amino acids causes translocation of mTORC1 to lysosomes, an obligate step for activation. To examine the spatial and temporal dynamics of this translocation, we used live imaging of the mTORC1 component RAPTOR and a cell permeant fluorescent analogue of di-leucine methyl ester. Translocation to lysosomes is a transient event, occurring within 2 min of aa addition and peaking within 5 min. It is temporally coupled with fluorescent leucine appearance in lysosomes and is sustained in comparison to aa stimulation. Sestrin2 and the vacuolar ATPase are negative and positive regulators of mTORC1 activity in our experimental system. Of note, phosphorylation of canonical mTORC1 targets is delayed compared to lysosomal translocation suggesting a dynamic and transient passage of mTORC1 from the lysosomal surface before targetting its substrates elsewhere. DOI: http://dx.doi.org/10.7554/eLife.19960.001 eLife Sciences Publications, Ltd 2016-10-11 /pmc/articles/PMC5059141/ /pubmed/27725083 http://dx.doi.org/10.7554/eLife.19960 Text en © 2016, Manifava et al http://creativecommons.org/licenses/by/4.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biochemistry
Manifava, Maria
Smith, Matthew
Rotondo, Sergio
Walker, Simon
Niewczas, Izabella
Zoncu, Roberto
Clark, Jonathan
Ktistakis, Nicholas T
Dynamics of mTORC1 activation in response to amino acids
title Dynamics of mTORC1 activation in response to amino acids
title_full Dynamics of mTORC1 activation in response to amino acids
title_fullStr Dynamics of mTORC1 activation in response to amino acids
title_full_unstemmed Dynamics of mTORC1 activation in response to amino acids
title_short Dynamics of mTORC1 activation in response to amino acids
title_sort dynamics of mtorc1 activation in response to amino acids
topic Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5059141/
https://www.ncbi.nlm.nih.gov/pubmed/27725083
http://dx.doi.org/10.7554/eLife.19960
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