Several cell stresses induce nuclear factor-kappaB (NF-κB) activation, which include irradiation, oxidation, and UV. Interestingly, serum-starving stress-induced NF-κB activation in COS cells, but not in COS-A717 cells. COS-A717 is a mutant cell line of COS cells that is defective of the NF-κB signaling pathway. We isolated genes with compensating activity for the NF-κB pathway and one gene encoded the G protein β2 (Gβ2). Gβ2 is one of the G protein-coupled receptor signaling effectors. In COS-A717 cells, Gβ2 expression is significantly reduced. In Gβ2 cDNA-transfected COS-A717 cells, the NF-κB activity was increased along with the recovery of Gβ2 expression. Furthermore, serum-starving stress induced the NF-κB activity in Gβ2-transfected COS-A717 cells. Consistently, the serum-starved COS cells with siRNA-reduced Gβ2 protein expression showed decreased NF-κB activity. These results indicate that Gβ2 is required for starvation-induced NF-κB activation and constitutive NF-κB activity. We propose that serum contains some molecule(s) that strongly inhibits NF-κB activation mediated through Gβ2 signaling.