Tan S, Somia N, Maher P, Schubert D
Cellular Neurobiology Laboratory, The Salk Institute for Biological Studies, La Jolla, California 92037, USA.
J Cell Biol. 2001 Mar 5;152(5):997-1006. doi: 10.1083/jcb.152.5.997.
Oxidative stress and highly specific decreases in glutathione (GSH) are associated with nerve cell death in Parkinson's disease. Using an experimental nerve cell model for oxidative stress and an expression cloning strategy, a gene involved in oxidative stress-induced programmed cell death was identified which both mediates the cell death program and regulates GSH levels. Two stress-resistant clones were isolated which contain antisense gene fragments of the translation initiation factor (eIF)2alpha and express a low amount of eIF2alpha. Sensitivity is restored when the clones are transfected with full-length eIF2alpha; transfection of wild-type cells with the truncated eIF2alpha gene confers resistance. The phosphorylation of eIF2alpha also results in resistance to oxidative stress. In wild-type cells, oxidative stress results in rapid GSH depletion, a large increase in peroxide levels, and an influx of Ca(2+). In contrast, the resistant clones maintain high GSH levels and show no elevation in peroxides or Ca(2+) when stressed, and the GSH synthetic enzyme gamma-glutamyl cysteine synthetase (gammaGCS) is elevated. The change in gammaGCS is regulated by a translational mechanism. Therefore, eIF2alpha is a critical regulatory factor in the response of nerve cells to oxidative stress and in the control of the major intracellular antioxidant, GSH, and may play a central role in the many neurodegenerative diseases associated with oxidative stress.
氧化应激和谷胱甘肽(GSH)的高度特异性降低与帕金森病中的神经细胞死亡有关。利用氧化应激的实验性神经细胞模型和表达克隆策略,鉴定出一个参与氧化应激诱导的程序性细胞死亡的基因,该基因既介导细胞死亡程序,又调节GSH水平。分离出两个抗应激克隆,它们含有翻译起始因子(eIF)2α的反义基因片段,并表达少量的eIF2α。当用全长eIF2α转染这些克隆时,敏感性得以恢复;用截短的eIF2α基因转染野生型细胞则赋予抗性。eIF2α的磷酸化也导致对氧化应激的抗性。在野生型细胞中,氧化应激导致GSH迅速耗竭、过氧化物水平大幅升高以及Ca(2+)内流。相反,抗应激克隆在受到应激时保持高GSH水平,过氧化物或Ca(2+)没有升高,并且GSH合成酶γ-谷氨酰半胱氨酸合成酶(γGCS)升高。γGCS的变化受翻译机制调控。因此,eIF2α是神经细胞对氧化应激反应以及主要细胞内抗氧化剂GSH控制中的关键调节因子,可能在许多与氧化应激相关的神经退行性疾病中起核心作用。