Activation of SAPK/JNKs in vitro.
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.
Chadee
Deborah N
DN
Department of Biological Sciences, University of Toledo, Toledo, OH, USA. deborah.chadee@utoledo.edu
Kyriakis
John M
JM
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods*; D055785:Gene Knockdown Techniques; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000276:immunology / Q000378:metabolism*; D020930:MAP Kinase Kinase Kinases / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D051379:Mice; D010750:Phosphoproteins / Q000276:immunology; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000235:genetics
D000906:Antibodies; D010750:Phosphoproteins; D034741:RNA, Small Interfering; D048031:JNK Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases