mTOR, AMPK, and GCN2 coordinate the adaptation of hepatic energy metabolic pathways in response to protein intake in the rat

Chotechuang N, Azzout-Marniche D, Bos C, Chaumontet C, Gausser/s N, Steiler T, Gaudichon C, Tome D. mTOR, AMPK, and GCN2 coordinate the adaptation of hepatic energy metabolic pathways in response to protein intake in the rat. Am J Physiol Endocrinol Metab 297: E 1313-E 1323, 2009. First published September 8, 2009; doi: 10.1152/ajpendo.91000.2008.--Three transduction pathways are involved in amino acid (AA) sensing in liver: mammalian target of rapamycin (roTOR), AMP-activated protein kinase (AMPK), and general control nondepressible kinase 2 (GCN2). However, no study has investigated the involvement of these signaling pathways in hepatic AA sensing. To address the question of liver AA sensing and signaling in response to a high-protein (HP) dietary supply, we investigated the changes in the phosphorylation state of hepatic mTOR (p-mTOR), AMPK[alpha] (p-AMPK[alpha]), and GCN2 (p-GCN2) by Western blotting. In rats fed a HP diet for 14 days, the hepatic p-AMPK[alpha] and p-GCN2 were lower (P < 0.001), and those of both the p-roTOR and eukaryotic initiation factor 4E-binding protein-1 phosphorylation (p-4E-BP1) were higher (P < 0.01) compared with rats receiving a normal protein (NP) diet. In hepatocytes in primary culture, high AA concentration decreased AMPK[alpha] phosphorylation whether insulin was present or not (P < 0.01). Either AAs or insulin can stimulate p-roTOR, but this is not sufficient for 4E-BP1 phosphorylation that requires both (P < 0.01). As expected, branchedchain AAs (BCAA) or leucine stimulated the phosphorylation of mTOR, but both insulin and BCAA or leucine are required for 4E-BP1 phosphorylation. GCN2 phosphorylation was reduced by both AAs and insulin(P < 0.01), suggesting for the first time that the translation inhibitor GCN2 senses not only the AA deficiency but also the AA increase in the liver. The present findings demonstrate that AAs and insulin exert a coordinated action on translation and involved mTOR, AMPK, and GCN2 transduction pathways. mammalian target of rapamycin; adenosine 5'-monophosphate-activated protein kinase; general control nondepressible kinase 2; transduction pathways; translation; high-protein diet doi: 10.1152/ajpendo.91000.2008