Cross talk between p38MAPK and ERK is mediated through MAPK-mediated protein phosphatase 2A catalytic subunit α and MAPK phosphatase-1 expression in human leukemia U937 cells

This study explores the signaling transduction cascade of ERK and p38 MAPK on regulating MAPK phosphatase-1 (MKP-1) and protein phosphatase 2A catalytic subunit α (PP2Acα) expression in caffeine-treated human leukemia U937 cells. Caffeine induced an increase in the intracellular Ca(2+) concentration and ROS generation leading to p38 MAPK activation and ERK inactivation, respectively. Caffeine treatment elicited MKP-1 down-regulation and PP2Acα up-regulation. The transfection of constitutively active MEK1 or pretreatment with SB202190 (p38 MAPK inhibitor) abolished the caffeine effect on MKP-1 and PP2Acα expression. Caffeine repressed ERK-mediated c-Fos phosphorylation but evoked p38 MAPK-mediated CREB phosphorylation. Knockdown of c-Fos and CREB by siRNA showed that c-Fos and CREB were responsible for MKP-1 and PP2Acα expression, respectively. Promoter and chromatin immunoprecipitating assay supported the role of c-Fos and CREB in regulating MKP-1 and PP2Acα expression. Moreover, transfection of dominant negative MKP-1 cDNA led to p38 MAPK activation and PP2Acα down-regulation in U937 cells, while PP2A inhibitor attenuated caffeine-induced ERK inactivation and MKP-1 down-regulation. Taken together, our data indicate that a reciprocal relationship between ERK-mediated MKP-1 expression and p38 MAPK-mediated PP2Acα expression crucially regulates ERK and p38 MAPK phosphorylation in U937 cells.