Diverse G protein-coupled receptors depend on G beta gamma heterodimers to promote cell polarization and survival via direct activation of PI3K gamma and potentially other effectors. These events involve full activation of AKT via its phosphorylation at Ser473, suggesting that mTORC2, the kinase that phosphorylates AKT at Ser473, is activated dowristream of G beta gamma. Thus, we tested the hypothesis that G beta gamma directly contributes to mTOR signaling. Here, we demonstrate that endogenous mTOR interacts with G beta gamma. Cell stimulation with serum modulates G beta gamma interaction with mTOR. The carboxyl terminal region of mTOR, expressed as a GST-fusion protein, including the serine/threonine kinase domain, binds G beta gamma heterodimers containing different G subunits, except G beta(4). Both, mTORC1 and mTORC2 complexes interact with G beta(1)gamma(2) which promotes phosphorylation of their respective substrates, p70S6K and ART. In addition, chronic treatment with rapamycin, a condition known to interfere with assembly of mTORC2, reduces the interaction between G beta gamma and mTOR and the phosphorylation of AKT; whereas overexpression of G alpha i interfered with the effect of G beta gamma as promoter of p70S6K and AKT phosphorylation. Altogether, our results suggest that G beta gamma positively regulates mTOR signaling via direct interactions and provide further support to emerging strategies based on the therapeutical potential of inhibiting different G beta gamma signaling interfaces. (C) 2014 Elsevier Inc. All rights reserved.