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2015 ; 10
(10
): 1767-1777
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Dynamic Visualization of mTORC1 Activity in Living Cells
#MMPMID25772363
Zhou X
; Clister TL
; Lowry PR
; Seldin MM
; Wong GW
; Zhang J
Cell Rep
2015[Mar]; 10
(10
): 1767-1777
PMID25772363
show ga
The mechanistic target of rapamycin complex 1 (mTORC1) senses diverse signals to
regulate cell growth and metabolism. It has become increasingly clear that mTORC1
activity is regulated in time and space inside the cell, but direct interrogation
of such spatiotemporal regulation is challenging. Here, we describe a genetically
encoded mTORC1 activity reporter (TORCAR) that exhibits a change in FRET in
response to phosphorylation by mTORC1. Co-imaging mTORC1 activity and calcium
dynamics revealed that a growth-factor-induced calcium transient contributes to
mTORC1 activity. Dynamic activity maps generated with the use of subcellularly
targeted TORCAR uncovered mTORC1 activity not only in cytosol and at the lysosome
but also in the nucleus and at the plasma membrane. Furthermore, a wide
distribution of activities was observed upon growth factor stimulation, whereas
leucine ester, an amino acid surrogate, induces more compartmentalized activities
at the lysosome and in the nucleus. Thus, mTORC1 activities are spatiotemporally
regulated in a signal-specific manner.