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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Bibliographic Details
Published in:eLife 2016-10, Vol.5
Main Authors: Manifava, Maria, Smith, Matthew, Rotondo, Sergio, Walker, Simon, Niewczas, Izabella, Zoncu, Roberto, Clark, Jonathan, Ktistakis, Nicholas T
Format: Article
Language:English
Subjects:
Adenosine triphosphatase
Amino acids
Amino Acids - metabolism
Autophagy
Biochemistry
Cell Biology
HEK293 Cells
Humans
Intravital Microscopy
Kinases
Leucine
Localization
Lysosomes
Lysosomes - metabolism
Mechanistic Target of Rapamycin Complex 1 - metabolism
mtor
Nuclear Proteins - metabolism
Nutrient interactions
Phosphorylation
Properties
Protein research
Protein synthesis
Protein Transport
Proteins
Regulatory-Associated Protein of mTOR - analysis
Sensors
signaling
Spatio-Temporal Analysis
Translocations (Genetics)
Vacuolar Proton-Translocating ATPases - metabolism
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Summary: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.
ISSN:2050-084X
2050-084X
DOI:10.7554/elife.19960