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.
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References
Kyriakis, J. M., and Avruch, J. (2001) Mammalian mitogen-activated protein kinase signal transduction pathways activated by stress and inflammation. Physiol Rev 81, 807–69.
Lei, K., and Davis, R. J. (2003) JNK phosphorylation of Bim-related members of the Bcl2 family induces Bax-dependent apoptosis. Proc Natl Acad Sci U S A 100, 2432–7.
Sharrocks, A. D., Yang, S. H., and Galanis, A. (2000) Docking domains and substrate specificity determination for MAP kinases. Trends Biochem Sci 25, 448–53.
Yoshioka, K. (2004) Scaffold proteins in mammalian MAP kinase cascades. J Biochem 135, 657–61.
Morrison, D. K., and Davis, R. J. (2003) Regulation of MAP kinase signaling modules by scaffold proteins in mammals. Annu Rev Cell Dev Biol 19, 91–118.
Chadee, D. N., and Kyriakis, J. M. (2004) MLK3 is required for mitogen activation of B-Raf, ERK and cell. Nat Cell Biol 6, 770–6.
Zhong, J., and Kyriakis, J. M. (2007) Dissection of a signaling pathway by which pathogen-associated molecular patterns recruit the JNK and p38 MAPKs and trigger cytokine release. J Biol Chem 282, 24246–54.
Chadee, D. N., Yuasa, T., and Kyriakis, J. M. (2002) Direct activation of mitogen-activated protein kinase kinase kinase MEKK1 by the Ste20p homologue GCK and the adapter protein TRAF2. Mol Cell Biol 22, 737–49.
Hibi, M., Lin, A., Smeal, T., Minden, A., and Karin, M. (1993) Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev 7, 2135–48.
Derijard, B., Hibi, M., Wu, I. H., Barrett, T., Su, B., Deng, T., Karin, M., and Davis, R. J. (1994) JNK1: a protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell 76, 1025–37.
Kyriakis, J. M., Banerjee, P., Nikolakaki, E., Dai, T., Rubie, E. A., Ahmad, M. F., Avruch, J., and Woodgett, J. R. (1994) The stress-activated protein kinase subfamily of c-Jun kinases. Nature 369, 156–60.
Kyriakis, J. M., Woodgett, J. R., and Avruch, J. (1995) The stress-activated protein kinases. A novel ERK subfamily responsive to cellular stress and inflammatory cytokines. Ann N Y Acad Sci 766, 303–19.
Galcheva-Gargova, Z., Derijard, B., Wu, I. H., and Davis, R. J. (1994) An osmosensing signal transduction pathway in mammalian cells. Science 265, 806–8.
Makkinje, A., Quinn, D. A., Chen, A., Cadilla, C. L., Force, T., Bonventre, J. V., and Kyriakis, J. M. (2000) Gene 33/Mig-6, a transcriptionally inducible adapter protein that binds GTP-Cdc42 and activates SAPK/JNK. A potential marker transcript for chronic pathologic conditions, such as diabetic nephropathy. Possible role in the response to persistent stress. J Biol Chem 275, 17838–47.
Shapiro, P. S., Evans, J. N., Davis, R. J., and Posada, J. A. (1996) The seven transmembrane-spanning receptors for endothelin and thrombin cause proliferation of airway smooth muscle cells and activation of the extracellular regulated kinase and c-Jun NH2-terminal kinase groups of mitogen-activated protein kinases. J Biol Chem 271, 5750–4.
Pandey, P., Raingeaud, J., Kaneki, M., Weichselbaum, R., Davis, R. J., Kufe, D., and Kharbanda, S. (1996) Activation of p38 mitogen-activated protein kinase by c-Abl dependent and -independent mechanisms. J Biol Chem 271, 23775–9.
Liu, H., Nishitoh, H., Ichijo, H., and Kyriakis, J. M. (2000) Activation of apoptosis signal-regulating kinase 1 (ASK1) by tumor necrosis factor receptor-associated factor 2 requires prior dissociation of the ASK1 inhibitor thioredoxin. Mol Cell Biol 20, 2198–208.
Pombo, C. M., Bonventre, J. V., Avruch, J., Woodgett, J. R., Kyriakis, J. M., and Force, T. (1994) The stress-activated protein kinases are major c-Jun amino-terminal kinases activated by ischemia and reperfusion. J Biol Chem 269, 26546–51.
Kyriakis, J. M., and Avruch, J. (1996) Sounding the alarm: protein kinase cascades activated by stress and inflammation. J Biol Chem 271, 24313–6.
Atfi, A., Djelloul, S., Chastre, E., Davis, R., and Gespach, C. (1997) Evidence for a role of Rho-like GTPases and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK) in transforming growth factor beta-mediated signaling. J Biol Chem 272, 1429–32.
Zhong, J., and Kyriakis, J. M. (2004) Germinal center kinase is required for optimal Jun N-terminal kinase activation by Toll-like receptor agonists and is regulated by the ubiquitin proteasome system and agonist-induced, TRAF6-dependent stabilization. Mol Cell Biol 20, 9165–75.
Zhong, J., Gavrilescu, L. C., Molnár, A., Murray, L., Garafalo, S., Kehrl, J. H., Simon, A. R., Van Etten, R. A., and Kyriakis, J. M. (2009) GCK is essential to systemic inflammation and pattern recognition receptor signaling to JNK and p38. Proc Natl Acad Sci U S A 106, 4372–7.
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Chadee, D.N., Kyriakis, J.M. (2010). Activation of SAPK/JNKs In Vitro. In: Seger, R. (eds) MAP Kinase Signaling Protocols. Methods in Molecular Biology, vol 661. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-795-2_3
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DOI: https://doi.org/10.1007/978-1-60761-795-2_3
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