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Glucagon Activates mTOR1/2 Signaling via an EPAC/Rap1 Signaling Axis in Hepatocyte Cultures

Activation of the protein kinase mTOR (mechanistic target of rapamycin) in complex 1/2 (mTORC1/2) in the liver is repressed during fasting and rapidly stimulated in response to feeding. The effect of a meal on hepatic mTORC1/2 is largely mediated by an increase in plasma levels of nutrients (i.e. amino acids) and growth factors (i.e. insulin). By contrast, fasting is associated with elevated plasma levels of glucagon, which is conventionally viewed as playing a counter‐regulatory role to insulin. However, glucagon and insulin secretion are both rapidly increased by consumption of a protein meal. In fact, amino acid‐induced glucagon secretion is many times greater than that seen with fasting. Herein we investigated the impact of insulin and glucagon on mTORC1/2 activation in rat H4IIE and human HepG2 cell cultures. As expected, insulin enhanced phosphorylation of the mTORC1 substrates p70S6K1 and 4E‐BP1. Surprisingly, the effect of glucagon on mTORC1 signaling was biphasic, wherein there was an acute increase in phosphorylation of p70S6K1 and 4E‐BP1 over the first hour of exposure, followed by latent suppression. Notably, the stimulatory effect of glucagon on mTORC1 was not additive with insulin, suggesting a convergent signaling axis. Glucagon‐induced mTORC1 activation was associated with enhanced cAMP levels and required activation of the glucagon receptor, PI3K/Akt, and exchange protein activated by cAMP (EPAC). An EPAC‐specific cAMP analog as well as PKA inhibition enhanced activation of mTORC1/2. EPAC acts as the guanine nucleotide exchange factor for the small GTPase Rap1, and in H4IIE cultures, transient expression of Rap1 was sufficient to stimulate activation of mTORC1/2. Finally, we found that glucagon addition to culture medium was sufficient to promote Rap1‐GTP loading in H4IIE cells. Overall, the findings support that glucagon elicits biphasic regulation of hepatic mTORC1/2 by an EPAC‐dependent increase in Rap1‐GTP which is followed by a repression of mTORC1.