title: The mammalian target of rapamycin (mTOR) pathway regulates mitochondrial oxygen consumption and oxidative capacity.
abstract: Metabolic rate and the subsequent production of reactive oxygen species are thought to contribute to the rate of aging in a wide range of species. The target of rapamycin (TOR) is a well conserved serine/threonine kinase that regulates cell growth in response to nutrient status. Here we demonstrate that in mammalian cells the mammalian TOR (mTOR) pathway plays a significant role in determining both resting oxygen consumption and oxidative capacity. In particular, we demonstrate that the level of complex formation between mTOR and one of its known protein partners, raptor, correlated with overall mitochondrial activity. Disruption of this complex following treatment with the mTOR pharmacological inhibitor rapamycin lowered mitochondrial membrane potential, oxygen consumption, and ATP synthetic capacity. Subcellular fractionation revealed that mTOR as well as mTOR-raptor complexes can be purified in the mitochondrial fraction. Using two-dimensional difference gel electrophoresis, we further demonstrated that inhibiting mTOR with rapamycin resulted in a dramatic alteration in the mitochondrial phosphoproteome. RNA interference-mediated knockdown of TSC2, p70 S6 kinase (S6K1), raptor, or rictor demonstrates that mTOR regulates mitochondrial activity independently of its previously identified cellular targets. Finally we demonstrate that mTOR activity may play an important role in determining the relative balance between mitochondrial and non-mitochondrial sources of ATP generation. These results may provide insight into recent observations linking the TOR pathway to life span regulation of lower organisms.
authors.affiliation: Cardiology Branch, Laboratory of Cardiac Energetics, Flow Cytometry Core Facility, NHLBI, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: Stefan M
authors.initials: SM
authors.lastname: Schieke
authors.forename: Darci
authors.initials: D
authors.lastname: Phillips
authors.forename: J Philip
authors.initials: JP
authors.lastname: McCoy
authors.forename: Angel M
authors.initials: AM
authors.lastname: Aponte
authors.forename: Rong-Fong
authors.initials: RF
authors.lastname: Shen
authors.forename: Robert S
authors.initials: RS
authors.lastname: Balaban
authors.forename: Toren
authors.initials: T
authors.lastname: Finkel
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D008564:Membrane Potentials; D008928:Mitochondria / Q000378:metabolism; D008954:Models, Biological; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption; D010766:Phosphorylation; D011494:Protein Kinases / Q000737:chemistry*; D040901:Proteomics / Q000379:methods; D034622:RNA Interference; D013347:Subcellular Fractions / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases
chemical_list: D000255:Adenosine Triphosphate; D011494:Protein Kinases; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D010100:Oxygen