Sequential Activation of Phosphatidylinositol 3-Kinase, βPix, Rac1, and Nox1 in Growth Factor-Induced Production of H2O2

The generation of reactive oxygen species (ROS) in cells stimulated with growth factors requires the activation of phosphatidylinositol 3-kinase (PI3K) and the Rac protein.We report here that the COOHterminal region of Nox1, a protein related to gp91 phox (Nox2) of phagocytic cells, is constitutively associated with ␤Pix, a guanine nucleotide exchange factor for Rac.Both growth factor-induced ROS production and Rac1 activation were completely blocked in cells depleted of ␤Pix by RNA interference.Rac1 was also shown to bind to the COOH-terminal region of Nox1 in a growth factor-dependent manner.Moreover, the depletion of Nox1 by RNA interference inhibited growth factor-induced ROS generation.These results suggest that ROS production in growth factor-stimulated cells is mediated by the sequential activation of PI3K, ␤Pix, and Rac1, which then binds to Nox1 to stimulate its NADPH oxidase activity.