DISTINCT EFFECTS OF GLUTATHIONE DISULFIDE ON THE NUCLEAR TRANSCRIPTION FACTORS KAPPA-B AND THE ACTIVATOR PROTEIN-1

Oxidative conditions potentiate the activation of the nuclear transcri ption factor kappa B (NF kappa B) and the activator protein-1 (AP-1) i n intact cells, but inhibit their DNA binding activity in vitro. We no w show that both the activation of NF kappa B and the inhibition of it s DNA binding activity is modulated in intact cells by the physiologic al oxidant glutathione disulphide (GSSG). NF kappa B activation in hum an T lineage cells (Molt-4) by 12-O-tetradecanoyl-phorbol 13-acetate w as inhibited by dithiothreitol, and this was partly reversed by the gl utathione reductase inhibitor 1,3-bis(2-chloroethyl)-1-nitrosourea (BC NU) or by hydrogen peroxide, indicating that GSSG may be required for NF kappa B activation. These effects of BCNU and hydrogen peroxide wer e not seen in glutathione-depleted cells. However, NF kappa B and AP-1 activation were potentiated by dithiothreitol if added to cell cultur es Ih after the phorbol ester, indicating that a shift of redox condit ions may support optimal oxidative activation with minimal inhibition of DNA binding. The elevation of intracellular GSSG levels by BCNU bef ore stimulation suppressed the chloramphenicol acetyltransferase expre ssion dependent on NF kappa B but increased that dependent on AP-1. Th is selective suppression of NF kappa B was also demonstrable by electr ophoretic mobility shift assays. In vitro, GSSG inhibited the DNA bind ing activity of NF kappa B more effectively than that of AP-1, while A P-1 was inhibited more effectively by oxidized thioredoxin.