Differential induction of nuclear NF-kappa B by protein phosphatase inhibitors in primary and transformed human cells. Requirement for both oxidation and phosphorylation in nuclear translocation.

Phosphoseryl/threonyl protein phosphatase inhibitors, viz. okadaic acid and calyculin-A, failed to induce nuclear factor-kappa B (NF-kappa B) nuclear translocation in several primary human cells although a marked and rapid induction was observed in their simian virus 40 transformed counterparts. Inability to induce NF-kappa B cannot be due to a non-activatable system since NF-kappa B was strongly activated by tumor necrosis factor in all the five primary cell types tested. It is also unlikely that the differential induction was due to differential sensitivity of primary and transformed cells to phosphatase inhibitors as the intracellular phosphatase activities of both cell types were equally inhibited by these inhibitors. However, pretreatment with hydrogen peroxide or buthionine sulfoximine, chemicals known to directly or indirectly elevate the intracellular free-radical levels, enabled okadaic acid to induce nuclear translocation of NF-kappa B in primary cells. Conversely cysteine, an antioxidant and precursor of the free radical scavenger, glutathione, inhibited the induction of NF-kappa B by tumor necrosis factor in primary cells, and by okadaic acid or tumor necrosis factor in transformed cells. These data, taken together, suggest that free radical-dependent oxidation and protein phosphorylation are not independent modes of NF-kappa B induction, but are both required for the release of NF-kappa B from I kappa B. Furthermore, the differential induction of NF-kappa B nuclear translocation by okadaic acid in primary and transformed human cells, reported herein, reflects intrinsic differences in the intracellular oxidative state between the two groups of cells. The induction of NF-kappa B by tumor necrosis factor in primary cells suggests that this cytokine fulfills the requirement for oxidation, possibly by inducing the production of free radicals.