Honda A, Nogami M, Yokozeki T, Yamazaki M, Nakamura H, Watanabe H, Kawamoto K, Nakayama K, Morris A J, Frohman M A, Kanaho Y
Department of Biological Information, Tokyo Institute of Technology, Yokohama, Japan.
Cell. 1999 Nov 24;99(5):521-32. doi: 10.1016/s0092-8674(00)81540-8.
Synthesis of phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], a signaling phospholipid, is primarily carried out by phosphatidylinositol 4-phosphate 5-kinase [PI(4)P5K], which has been reported to be regulated by RhoA and Rac1. Unexpectedly, we find that the GTPgammaS-dependent activator of PI(4)P5Kalpha is the small G protein ADP-ribosylation factor (ARF) and that the activation strictly requires phosphatidic acid, the product of phospholipase D (PLD). In vivo, ARF6, but not ARF1 or ARF5, spatially coincides with PI(4)P5Kalpha. This colocalization occurs in ruffling membranes formed upon AIF4 and EGF stimulation and is blocked by dominant-negative ARF6. PLD2 similarly translocates to the ruffles, as does the PH domain of phospholipase Cdelta1, indicating locally elevated PI(4,5)P2. Thus, PI(4)P5Kalpha is a downstream effector of ARF6 and when ARF6 is activated by agonist stimulation, it triggers recruitment of a diverse but interactive set of signaling molecules into sites of active cytoskeletal and membrane rearrangement.
磷脂酰肌醇4,5-二磷酸[PI(4,5)P2]是一种信号磷脂,其合成主要由磷脂酰肌醇4-磷酸5-激酶[PI(4)P5K]完成,据报道该激酶受RhoA和Rac1调节。出乎意料的是,我们发现PI(4)P5Kα的GTPγS依赖性激活剂是小G蛋白ADP-核糖基化因子(ARF),且这种激活严格需要磷脂酶D(PLD)的产物磷脂酸。在体内,ARF6而非ARF1或ARF5在空间上与PI(4)P5Kα重合。这种共定位发生在AIF4和表皮生长因子(EGF)刺激后形成的皱膜中,并被显性负性ARF6阻断。PLD2同样易位至皱膜,磷脂酶Cδ1的PH结构域也是如此,这表明局部PI(4,5)P2水平升高。因此,PI(4)P5Kα是ARF6的下游效应器,当ARF6被激动剂刺激激活时,它会触发多种但相互作用的信号分子募集到活跃的细胞骨架和膜重排位点。