title: Control of Ser2448 phosphorylation in the mammalian target of rapamycin by insulin and skeletal muscle load.
abstract: We have investigated the effects of insulin, amino acids, and the degree of muscle loading on the phosphorylation of Ser(2448), a site in the mammalian target of rapamycin (mTOR) phosphorylated by protein kinase B (PKB) in vitro. Phosphorylation was assessed by immunoblotting with a phosphospecific antibody (anti-Ser(P)(2448)) and with mTAb1, an activating antibody whose binding is inhibited by phosphorylation in the region of mTOR that contains Ser(2448). Incubating rat diaphragm muscles with insulin increased Ser(2448) phosphorylation but did not change the total amount of mTOR. Insulin, but not amino acids, activated PKB, as evidenced by increased phosphorylation of both Ser(308) and Thr(473) in the kinase. Ser(2448) phosphorylation was also modulated by muscle-loading. Overloading the rat plantaris muscle by synergist muscle ablation, which promotes hypertrophy of the plantaris muscle, increased Ser(2448) phosphorylation. In contrast, unloading the gastrocnemius muscle by hindlimb suspension, which promotes atrophy of the muscle, decreased Ser(2448) phosphorylation, an effect that was fully reversible. Neither overloading nor hindlimb suspension significantly changed the total amount of mTOR. In summary, our results demonstrate that atrophy and hypertrophy of skeletal muscle are associated with decreases and increases in Ser(2448) phosphorylation, suggesting that modulation of this site may have an important role in the control of protein synthesis.
authors.affiliation: Department of Pharmacology, University of Virginia Health System, Charlottesville, Virginia 22908-0735, USA.
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Reynolds
authors.forename: Sue C
authors.initials: SC
authors.lastname: Bodine
authors.forename: John C
authors.initials: JC
authors.lastname: Lawrence
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D003964:Diaphragm / Q000201:enzymology / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000276:immunology / Q000378:metabolism; D007328:Insulin / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000502:physiology*; D010766:Phosphorylation; D014176:Protein Biosynthesis; D011494:Protein Kinases / Q000378:metabolism / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism; D012694:Serine / Q000378:metabolism*; D020123:Sirolimus / Q000378:metabolism*; D058570:TOR Serine-Threonine Kinases; D016474:Weight-Bearing
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000596:Amino Acids; D015699:HIV Envelope Protein gp120; D007328:Insulin; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; C486300:TAB1 protein, human; D012694:Serine; D011494:Protein Kinases; C546842:MTOR protein, human; C546845:mTOR protein, rat; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D058570:TOR Serine-Threonine Kinases; D020123:Sirolimus