Activation of SAPK/JNKs in vitro.

Abstract

The stress-activated protein kinase/c-jun N-terminal kinases (SAPK/JNKs) are mitogen-activated protein kinases (MAPKs) that are activated by stressful and inflammatory stimuli and regulate cellular responses such as proliferation, differentiation, and apoptosis. The SAPK/JNKs are phosphorylated and activated by the MAP kinase kinases (MAP2Ks), SEK1/MKK4 and MKK7. These MAP2Ks are phosphorylated and activated by upstream stress-activated MAPK kinase kinases (MAP3Ks). Upon activation, SAPK/JNKs translocate to the nucleus and phosphorylate transcription factors, ultimately resulting in the modulation of gene expression. We have analyzed the activation of SAPK/JNK and stress-activated MAP3Ks using in vitro kinase assays. In addition, we have studied the role of different MAP3Ks in SAPK/JNK signaling by silencing specific MAP3K expression with RNAi and then analyzing the effect on activation of SAPK/JNKs and other MAPKs.