The p38 MAP kinase inhibitor SB203580 enhances nuclear factor-kappa B transcriptional activity by a non-specific effect upon the ERK pathway

1 In the present study we investigated a possible role for the p38 mitogen- activated protein (MAP) kinase pathway in mediating nuclear factor-kappa B (NF-kappa B) transcriptional activity in the erythroleukaemic cell line TF- 1. 2 TF-1 cells stimulated with the phosphatase inhibitor okadaic acid (OA) de monstrated enhanced NF-kappa B and GAL4p65-regulated transcriptional activi ty which was associated with elevated p38 phosphorylation. However, pretrea tment with the p38 MAPK specific inhibitor SB203580 (1 mu M) or overexpress ion of kinase-deficient mutants of MKK3 or MKK6 did not affect OA-enhanced NF-kappa B transcriptional potency, as determined in transient transfection assays. In fact, 5 and 10 mu M SB203580 enhanced rather than inhibited NF- kappa B-mediated promoter activity by 2 fold, which was independent of phos phorylation of the p65 subunit. 3 The SB203580-mediated increase in NF-kappa B transcriptional activity was associated with enhanced phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK), but not p38 kinase. 4 Overexpression of kinase-deficient mutants belonging to the ERK1/2, JNK, and p38 pathways showed that only dominant-negative Raf-l abrogated SB20358 0-enhanced NF-kappa B activity. This would implicate the involvement of the ERK1/2 pathway in the enhancing effects of SB203580 on NF-kappa B-mediated gene transcription. 5 This study demonstrates that the p38 MAP kinase pathway is not involved i n the OA-induced activation of NF-kappa B. SB203580 at higher concentration s activates the ERK pathway, which subsequently enhances NF-kappa B transcr iptional activity.