Dynamic Visualization of mTORC1 Activity in Living Cells

Xin Zhou, Terri L. Clister, Pamela R. Lowry, Marcus M. Seldin, G. William Wong, Jin Zhang

Research output: Contribution to journalArticle

Abstract

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 andspace inside the cell, but direct interrogation ofsuch 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.

Original languageEnglish (US)
Pages (from-to)1767-1777
Number of pages11
JournalCell Reports
Volume10
Issue number10
DOIs
StatePublished - Mar 17 2015

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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    Zhou, X., Clister, T. L., Lowry, P. R., Seldin, M. M., Wong, G. W., & Zhang, J. (2015). Dynamic Visualization of mTORC1 Activity in Living Cells. Cell Reports, 10(10), 1767-1777. https://doi.org/10.1016/j.celrep.2015.02.031