Adenovirus type 5 E1A protein (E1A) associates with anti-tumor activities by reversing the transformed phenotype, inhibiting metastasis, and inducing apoptosis. We have previously identified that E1A suppresses the expression of Axl, a transforming tyrosine kinase and that Axl-Gas6 receptor-ligand interaction prevents E1A transfectants from apoptosis induced by serum deprivation. To determine how the Axl-Gas6 interaction prevents E1A-mediated apoptosis, we analysed the expression of anti-apoptotic molecules and found that the activated form of Akt was suppressed in the E1A transfectant ip 1-E1A and that Gas6 was able to activate Akt in ip 1-E1A cells reexpressing Axl (ip 1-E1A-Axl). To determine whether activated Akt is required to prevent E1A-mediated apoptosis, ip 1-E1A-Axl cells were treated with the phosphatidylinositol-3'-OH kinase inhibitor wortmannin or transfected with a dominant negative Akt mutant. In both cases, Gas6 no longer protected the cells from serum deprivation-induced apoptosis. Thus, we conclude that activated Akt is required for Axl-Gas6 signaling to prevent E1A-mediated apoptosis induced by serum deprivation. Downstream molecules of Akt, including NF-kappaB, Fas ligand, and BAD were examined, among which phosphorylation of BAD by Axl-Gas6 signaling is associated with the anti-apoptotic activity of Akt in our study.