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Protein phosphorylation is involved in the induction of nitric oxide synthase II (NOS II, iNOS) in several types of animal cells. Here we have investigated the possible involvement of major protein kinases in the induction of NOS II expression in human DLD‐1 cells. In DLD‐1 cells, interferon‐γ alone induced a submaximal NOS II expression; a cytokine mixture consisting of interferon‐γ, tumour necrosis factor‐α and interleukin‐1β produced maximal NOS II induction. Activators of protein kinase A (forskolin, 8‐dibutyryl‐cyclic AMP), of protein kinase C (tetradecanoylphorbol‐13‐acetate), and of protein kinase G (8‐bromo cyclic GMP) did not induce NOS II mRNA by themselves, nor did they alter NOS II mRNA induction in response to cytokines. Inhibitors of protein kinase A (compound H89), of protein kinase C (bisindolylmaleimide, chelerythrine or staurosporine), of phosphatidylinositol 3‐kinase (wortmannin), of p38 mitogen‐activated protein kinase (compound SB 203580) and of extracellular signal‐regulated kinase (compound PD 98059) also had no influence on basal or cytokine‐induced NOS II mRNA expression. Immunoprecipitation kinase assays showed no activation of extracellular signal‐regulated kinase or p38 mitogen‐activated protein kinase in cytokine‐incubated DLD‐1 cells. The c‐Jun NH2‐terminal kinase was activated by cytokines, but the most efficacious cytokine was tumour necrosis factor‐α which did not induce NOS II by itself. In contrast, the protein tyrosine kinase inhibitor tyrphostin B42 (a specific inhibitor of interferon‐γ‐activated janus kinase 2) and the protein tyrosine kinase inhibitor tyrphostin A25 both reduced CM‐induced NOS II mRNA expression in a concentration‐dependent manner. These results suggest that activation of NOS II expression in DLD‐1 cells is independent of the activities of protein kinases A, C and G, phosphatidylinositol 3‐kinase, extracellular signal regulated kinase and p38 mitogen‐activated protein kinase, but seems to require protein tyrosine kinase activity, especially the interferon‐γ‐activated janus kinase 2. British Journal of Pharmacology (1998) 123, 1716–1722; doi:10.1038/sj.bjp.0701782
British Journal of Pharmacology – Wiley
Published: Apr 1, 1998
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