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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...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2016
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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 |
format | Online Article Text |
id | pubmed-5059141 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
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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