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PNAS 106 (52): 22229-22232

Copyright © 2009 by the National Academy of Sciences.

Direct control of mitochondrial function by mTOR

Arvind Ramanathan Stuart L. Schreiber1

Chemical Biology Program, Broad Institute of Harvard and Massachusetts Institute of Technology, 7 Cambridge Center, Cambridge, MA 02142


Figure 1
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Fig. 1.. Effect of rapamycin on mitochondria and glycolysis. (A) Jurkat cells were treated with 100 nM rapamycin in the presence or absence of the transcription inhibitor actinomycin D for 45 min. (B) Levels of maximally uncoupled respiration were measured after treatment with rapamycin or vehicle for 30 min, followed by treatment with 1 µm FCCP for 15 min. (C) Levels of lactic acid in the extracellular medium were measured after cells were treated with rapamycin for 45 min. *, P < .05. All experiments were performed in biological triplicate.

 

Figure 2
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Fig. 2.. mTOR affects mitochondrial metabolism via a complex with Bcl-xl. (A) Bcl-xl interacts with mTOR and VDAC1. Interaction of Bcl-xl with mTOR was decreased by treatment with rapamycin. (B) An in vitro kinase assay was performed using recombinant purified mTOR and Bcl-xl proteins and {gamma}P32-labeled ATP. Bcl-xl was detected using a Ponceau stain, and the phosphorylated serine 62 on Bcl-xl was detected using an antibody. (C) mTOR-mediated phosphorylation of Bcl-xl was monitored using recombinant, purified Bcl-xl protein at indicated concentrations, using an ATP-coupled luminescence assay, with 100 ng of mTOR protein used per reaction. (D) Jurkat cells were treated with rapamycin and ABT-263 at indicated concentrations, and oxygen consumption was measured. (E) Jurkat cells overexpressing either GFP or Bcl-xl were treated with DMSO vehicle or rapamycin, and oxygen consumption was measured. *, P < .05. All experiments were preformed in biological triplicate.

 


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