Details

Autor(en) / Beteiligte

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

Titel

Dynamic Visualization of mTORC1 Activity in Living Cells

Ist Teil von

• Cell reports (Cambridge), 2015-03, Vol.10 (10), p.1767-1777

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2015

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• 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. [Display omitted] •A genetically encoded mTORC1 activity reporter has been developed and characterized•A transient intracellular Ca2+ increase contributes to PDGF-induced mTORC1 activity•Akt plays a key role in mediating growth factor-induced lysosomal mTORC1 activity•mTORC1 is intricately regulated in a signal- and location-specific manner mTORC1 integrates diverse signals to regulate cell growth and metabolism, yet its activity has not been systematically characterized. Zhou et al. develop a genetically encoded mTORC1 activity reporter, uncover a signal-specific activity map of mTORC1, and demonstrate a wide distribution of mTORC1 activity in the plasma membrane, cytosol, lysosome, and nucleus.