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)2α and express a low amount of eIF2α. Sensitivity is restored when the clones are transfected with full-length eIF2α; transfection of wild-type cells with the truncated eIF2α gene confers resistance. The phosphorylation of eIF2α 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 Ca²⁺. In contrast, the resistant clones maintain high GSH levels and show no elevation in peroxides or Ca²⁺ when stressed, and the GSH synthetic enzyme γ-glutamyl cysteine synthetase (γGCS) is elevated. The change in γGCS is regulated by a translational mechanism. Therefore, eIF2α 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.