Complex interactions between the angiotensin II type 1 receptor, the epidermal growth factor receptor and...

Transactivation of the epidermal growth factor receptor (EGFR) by the angiotensin II (AngII) type 1 (AT₁) receptor is involved in AT₁ receptor-dependent growth effects and cardiovascular pathologies, however the mechanisms underpinning this transactivation are yet to be fully elucidated. Recently, a potential intermediate of this process was identified following the discovery that a kinase called TRIO was involved in AngII/AT₁ receptor-mediated transactivation of EGFR. To investigate the mechanisms by which TRIO acts as an intermediate in AngII/AT₁ receptor-mediated EGFR transactivation we used bioluminescence resonance energy transfer (BRET) assays to investigate proximity between the AT₁ receptor, EGFR, TRIO and other proteins of interest. We found that AngII/AT₁ receptor activation caused a Gα_q-dependent increase in proximity of TRIO with Gγ₂ and the AT₁-EGFR heteromer, as well as trafficking of TRIO towards the Kras plasma membrane marker and into early, late and recycling endosomes. In contrast, we found that AngII/AT₁ receptor activation caused a Gα_q-independent increase in proximity of TRIO with Grb2, GRK2 and PKCζ, as well as trafficking of TRIO up to the plasma membrane from the Golgi. Furthermore, we confirmed the proximity between the AT₁ receptor and the EGFR using the Receptor-Heteromer Investigation Technology, which showed AngII-induced recruitment of Grb2, GRK2, PKCζ, Gγ₂ and TRIO to the EGFR upon AT₁ coexpression. In summary, our results provide further evidence for the existence of the AT₁-EGFR heteromer and reveal potential mechanisms by which TRIO contributes to the transactivation process.