Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
J Biol Chem. 2011 Aug 26;286(34):29758-70. doi: 10.1074/jbc.M111.263103. Epub 2011 Jun 28.
Phosphoinositide 3-kinase (PI3K) regulates cell polarity and migration by generating phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P(3)) at the leading edge of migrating cells. The serine-threonine protein kinase Akt binds to PI(3,4,5)P(3), resulting in its activation. Active Akt promotes spatially regulated actin cytoskeletal remodeling and thereby directed cell migration. The inositol polyphosphate 5-phosphatases (5-ptases) degrade PI(3,4,5)P(3) to form PI(3,4)P(2), which leads to diminished Akt activation. Several 5-ptases, including SKIP and SHIP2, inhibit actin cytoskeletal reorganization by opposing PI3K/Akt signaling. In this current study, we identify a molecular co-chaperone termed silencer of death domains (SODD/BAG4) that forms a complex with several 5-ptase family members, including SKIP, SHIP1, and SHIP2. The interaction between SODD and SKIP exerts an inhibitory effect on SKIP PI(3,4,5)P(3) 5-ptase catalytic activity and consequently enhances the recruitment of PI(3,4,5)P(3)-effectors to the plasma membrane. In contrast, SODD(-/-) mouse embryonic fibroblasts exhibit reduced Akt-Ser(473) and -Thr(308) phosphorylation following EGF stimulation, associated with increased SKIP PI(3,4,5)P(3)-5-ptase activity. SODD(-/-) mouse embryonic fibroblasts exhibit decreased EGF-stimulated F-actin stress fibers, lamellipodia, and focal adhesion complexity, a phenotype that is rescued by the expression of constitutively active Akt1. Furthermore, reduced cell migration was observed in SODD(-/-) macrophages, which express the three 5-ptases shown to interact with SODD (SKIP, SHIP1, and SHIP2). Therefore, this study identifies SODD as a novel regulator of PI3K/Akt signaling to the actin cytoskeleton.
磷酸肌醇 3-激酶(PI3K)通过在迁移细胞的前缘生成磷脂酰肌醇 3,4,5-三磷酸(PI(3,4,5)P(3))来调节细胞极性和迁移。丝氨酸-苏氨酸蛋白激酶 Akt 与 PI(3,4,5)P(3)结合,导致其激活。活性 Akt 促进空间调节肌动蛋白细胞骨架重塑,从而促进细胞定向迁移。肌醇多磷酸 5-磷酸酶(5-ptases)将 PI(3,4,5)P(3)降解为 PI(3,4)P(2),从而导致 Akt 激活减少。几种 5-ptases,包括 SKIP 和 SHIP2,通过拮抗 PI3K/Akt 信号来抑制肌动蛋白细胞骨架重组。在本研究中,我们鉴定了一种称为死亡结构域沉默子(SODD/BAG4)的分子伴侣,它与包括 SKIP、SHIP1 和 SHIP2 在内的几种 5-ptase 家族成员形成复合物。SODD 与 SKIP 的相互作用对 SKIP PI(3,4,5)P(3)5-ptase 催化活性产生抑制作用,从而增强 PI(3,4,5)P(3)效应物向质膜的募集。相比之下,EGF 刺激后 SODD(-/-) 小鼠胚胎成纤维细胞的 Akt-Ser(473)和 -Thr(308)磷酸化减少,与 SKIP PI(3,4,5)P(3)-5-ptase 活性增加相关。SODD(-/-) 小鼠胚胎成纤维细胞表现出 EGF 刺激的 F-肌动蛋白应力纤维、片状伪足和焦点粘附复合物减少,该表型可通过表达组成型激活 Akt1 得到挽救。此外,在 SODD(-/-) 巨噬细胞中观察到细胞迁移减少,SODD(-/-) 巨噬细胞表达三种与 SODD(SKIP、SHIP1 和 SHIP2)相互作用的 5-ptases。因此,本研究确定 SODD 是 PI3K/Akt 信号向肌动蛋白细胞骨架的新型调节剂。