During feeding, increases in circulating pancreatic insulin inhibit hepatic glucose output through the activation of the Ser/Thr kinase AKT and subsequent phosphorylation of the forkhead transcription factor FOXO1 (refs 1–3). Under fasting conditions, FOXO1 increases gluconeogenic gene expression in concert with the cAMP responsive coactivator TORC2 (refs 4–8). In response to pancreatic glucagon, TORC2 is de-phosphorylated at Ser 171 and transported to the nucleus, in which it stimulates the gluconeogenic programme by binding to CREB. Here we show in mice that insulin inhibits gluconeogenic gene expression during re-feeding by promoting the phosphorylation and ubiquitin-dependent degradation of TORC2. Insulin disrupts TORC2 activity by induction of the Ser/Thr kinase SIK2, which we show here undergoes AKT2-mediated phosphorylation at Ser 358. Activated SIK2 in turn stimulated the Ser 171 phosphorylation and cytoplasmic translocation of TORC2. Phosphorylated TORC2 was degraded by the 26S proteasome during re-feeding through an association with COP1, a substrate receptor for an E3 ligase complex that promoted TORC2 ubiquitination at Lys 628. Because TORC2 protein levels and activity were increased in diabetes owing to a block in TORC2 phosphorylation, our results point to an important role for this pathway in the maintenance of glucose homeostasis.
We thank R. Crystal for Ad-CRE-Luc and Ad-Rsv-Luc reporters. We also thank J. Meisenhelder for help with two-dimensional tryptic mapping and L. Vera for mouse tail vein injections. This work was supported by NIH grants. R.D. is a recipient of a post-doctoral fellowship from the Fondation pour la Recherche Médicale and is supported by the Bettencourt Schuller Foundation. Y.L. is a Hillblom Foundation Fellow. S.-H.K. was supported by a grant of the Korea Health 21 R&D Project, Ministry of Health & Welfare, Republic of Korea. M.M. is supported by the Keickhefer Foundation. This research was also supported in part by the Foundation for Medical Research, Inc.
The file contains Supplementary Figures S1-S30 with Legends.