Autophagy modulates amino acid signaling network in myotubes: differential effects on mTORC1 pathway and the integrated stress response.
Induction of autophagy and the integrated stress response is important for amino acid homeostasis. It remains unknown whether the autophagy coregulates both mechanistic target of rapamycin complex 1 (mTORC1) signaling and the integrated stress response. In mouse C2C12 myotubes, we found that amino acid limitation induced autophagy and that the subsequent release of amino acid is required to sustain mTORC1 signaling. Inhibition of autophagy by bafilomycin A1 or chloroquine treatment during amino acid scarcity abolished mTORC1 signaling, an effect that could be rescued by inhibiting protein synthesis or amino acid supplementation, respectively. Autophagy is required to sustain the balance of both essential and nonessential amino acids during amino acid starvation, and it has a predominant role over the ubiquitin-proteasome system in the regulation of mTORC1. Inhibition of autophagy was found to activate the integrated stress response, as well as eukaryotic initiation factor 2α (eIF2α) and its target genes independent of amino acid availability. Conversely, autophagy induction via mTOR inhibition is sufficient to reduce eIF2α phosphorylation. Thus, autophagy protects the eIF2α-mediated stress response independent of amino acid supply in cultured myotubes. Our results showed that autophagy uniquely modulates mTORC1 and the integrated stress response in an amino acid-dependent and -independent manner, respectively.
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
Xinlei
X
Yu
Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore bchlongy@nus.edu.sg.
Yun Chau
YC
Long
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001343:Autophagy; D002460:Cell Line; D002738:Chloroquine / Q000737:chemistry; D015852:Eukaryotic Initiation Factor-2 / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006706:Homeostasis; D008247:Lysosomes / Q000378:metabolism; D018942:Macrolides / Q000737:chemistry; D000076222:Mechanistic Target of Rapamycin Complex 1; D051379:Mice; D046912:Multiprotein Complexes / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism; D010766:Phosphorylation; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D015398:Signal Transduction; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism
D000596:Amino Acids; D015852:Eukaryotic Initiation Factor-2; D018942:Macrolides; D046912:Multiprotein Complexes; D025801:Ubiquitin; D049452:Green Fluorescent Proteins; D002738:Chloroquine; C040929:bafilomycin A1; D000076222:Mechanistic Target of Rapamycin Complex 1; D058570:TOR Serine-Threonine Kinases; D046988:Proteasome Endopeptidase Complex
atrophy; lysosome; protein; proteolysis; skeletal muscle