Class III phosphoinositide 3-kinase--Beclin1 complex mediates the amino acid-dependent regulation of autophagy in C2C12 myotubes.
Increased proteolysis contributes to muscle atrophy that prevails in many diseases. Elucidating the signalling pathways responsible for this activation is of obvious clinical importance. Autophagy is a ubiquitous degradation process, induced by amino acid starvation, that delivers cytoplasmic components to lysosomes. Starvation markedly stimulates autophagy in myotubes, and the present studies investigate the mechanisms of this regulation. In C(2)C(12) myotubes incubated with serum growth factors, amino acid starvation stimulated autophagic proteolysis independently of p38 and p42/p44 mitogen-activated protein kinases, but in a PI3K (phosphoinositide 3-kinase)-dependent manner. Starvation, however, did not alter activities of class I and class II PI3Ks, and was not sufficient to affect major signalling proteins downstream from class I PI3K (glycogen synthase kinase, Akt/protein kinase B and protein S6). In contrast, starvation increased class III PI3K activity in whole-myotube extracts. In fact, this increase was most pronounced for a population of class III PI3K that coimmunoprecipitated with Beclin1/Apg6 protein, a major determinant in the initiation of autophagy. Stimulation of proteolysis was reproduced by feeding myotubes with synthetic dipalmitoyl-PtdIns3 P, the class III PI3K product. Conversely, protein transfection of anti-class III PI3K inhibitory antibody into starved myotubes inverted the induction of proteolysis. Therefore, independently of class I PI3K/Akt, protein S6 and mitogen-activated protein kinase pathways, amino acid starvation stimulates proteolysis in myotubes by regulating class III PI3K-Beclin1 autophagic complexes.
Tassa
Amina
A
Human Nutrition Research Center, Nutrition and Protein Metabolism Laboratory, INRA UR551, 63122 Ceyrat, France.
Roux
Marie Paule
MP
Attaix
Didier
D
Bechet
Daniel M
DM
D000596:Amino Acids / Q000502:physiology*; D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000235:genetics; D051017:Apoptosis Regulatory Proteins; D001343:Autophagy; D000071186:Beclin-1; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D046911:Macromolecular Substances; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000145:classification / Q000502:physiology*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011506:Proteins / Q000502:physiology*; D014162:Transfection; D000077191:Wortmannin
D000596:Amino Acids; D000730:Androstadienes; D000906:Antibodies; D051017:Apoptosis Regulatory Proteins; D000071186:Beclin-1; D004791:Enzyme Inhibitors; D046911:Macromolecular Substances; D020928:Mitogen-Activated Protein Kinases; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011506:Proteins; D000077191:Wortmannin; C115010:Becn1 protein, mouse