TY - JOUR T1 - Phosphatidylinositol 3-Kinase Regulates Nuclear Translocation of NF-E2-Related Factor 2 through Actin Rearrangement in Response to Oxidative Stress JF - Molecular Pharmacology JO - Mol Pharmacol SP - 1001 LP - 1010 DO - 10.1124/mol.62.5.1001 VL - 62 IS - 5 AU - Keon Wook Kang AU - Seung Jin Lee AU - Jeong Weon Park AU - Sang Geon Kim Y1 - 2002/11/01 UR - http://molpharm.aspetjournals.org/content/62/5/1001.abstract N2 - Expression of phase II detoxifying genes is regulated by NF-E2–related factor 2 (Nrf2)-mediated antioxidant response element (ARE) activation. We showed previously that phosphatidylinositol 3 (PI3)-kinase plays an essential role in ARE-mediated rGSTA2 induction by oxidative stress. In view of the fact that the signaling pathway of PI3-kinase controls microfilaments and translocation of actin-associated proteins, the current study was designed to investigate the PI3-kinase–mediated nuclear translocation of Nrf2 and the interaction of Nrf2 with actin.tert-Butylhydroquinone (t-BHQ) caused Nrf2 to translocate into the nucleus in H4IIE cells, which was prevented by pretreatment of the cells with PI3-kinase inhibitors (wortmannin/LY294002). t-BHQ relocalized Nrf2 in concert with changes in actin microfilament architecture, as visualized by superposition of immunochemically stained Nrf2 and fluorescent phalloidin-stained actin. Furthermore, t-BHQ increased the level of nuclear actin, coimmunoprecipitated with Nrf2, which returned to that of control by pretreatment of the cells with PI3-kinase inhibitors. Cytochalasin B, an actin disruptor, alone stimulated actin-mediated nuclear translocation of Nrf2 and induced rGSTA2. In contrast, phalloidin, an agent that prevents actin filaments from depolymerization, inhibited Nrf2 translocation and rGSTA2 induction by t-BHQ. Subcellular fractionation and immunoblot analyses allowed us to detect both 57- and 100-kDa Nrf2. Immunoblot and immunoprecipitation assays showed that the 100-kDa protein comprised both Nrf2 and actin. The present study demonstrates that the PI3-kinase signaling pathway regulates rearrangement of actin microfilaments in response to oxidative stress and that depolymerization of actin causes a complex of Nrf2 bound with actin to translocate into nucleus. ER -