Signaling-responsive MAP kinases (MAPKs) are key in mediating immune responses and are activated through the phosphorylation of a Thr-X-Tyr motif by upstream MAPK kinases. Here we show that T cells stimulated through the T cell receptor (TCR) used an alternative mechanism in which p38 was phosphorylated on Tyr323 and subsequently autophosphorylated residues Thr180 and Tyr182. This required the TCR-proximal tyrosine kinase Zap70 but not the adaptor protein LAT, which was required for activation of extracellular signal–regulated protein kinase MAPKs. TCR activation of p38 lacking Tyr323 was diminished, and blocking of p38 activity prevented p38 dual phosphorylation in normal T cells but not in B cells. Thus, phosphorylation of Tyr323 dependent on the tyrosine kinase Lck and mediated by Zap70 serves as an important mechanism for TCR activation of p38 in T cells.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Chang, L. & Karin, M. Mammalian MAP kinase signalling cascades. Nature 410, 37–40 (2001).
Dong, C., Davis, R.J. & Flavell, R.A. MAP kinases in the immune response. Annu. Rev. Immunol. 20, 55–72 (2002).
Rincon, M. et al. Conference highlight: do T cells care about the mitogen-activated protein kinase signalling pathways. Immunol. Cell Biol. 78, 166–175 (2000).
Rincon, M., Flavell, R.A. & Davis, R.A. The JNK and P38 MAP kinase signaling pathways in T cell-mediated immune responses. Free Radic. Biol. Med. 28, 1328–1337 (2000).
Rincon, M., Flavell, R.A. & Davis, R.J. Signal transduction by MAP kinases in T lymphocytes. Oncogene 20, 2490–2497 (2001).
Schaeffer, H.J. & Weber, M.J. Mitogen-activated protein kinases: specific messages from ubiquitous messengers. Mol. Cell. Biol. 19, 2435–2444 (1999).
Davis, R.J. Signal transduction by the JNK group of MAP kinases. Cell 103, 239–252 (2000).
Dong, C. et al. JNK is required for effector T-cell function but not for T-cell activation. Nature 405, 91–94 (2000).
Han, J. & Ulevitch, R.J. Emerging targets for anti-inflammatory therapy. Nat. Cell Biol. 1, E39–E40 (1999).
Rincon, M. MAP-kinase signaling pathways in T cells. Curr. Opin. Immunol. 13, 339–345 (2001).
Bellon, S., Fitzgibbon, M.J., Fox, T., Hsiao, H.M. & Wilson, K.P. The structure of phosphorylated p38γ is monomeric and reveals a conserved activation-loop conformation. Structure Fold. Des. 7, 1057–1065 (1999).
Canagarajah, B.J., Khokhlatchev, A., Cobb, M.H. & Goldsmith, E.J. Activation mechanism of the MAP kinase ERK2 by dual phosphorylation. Cell 90, 859–869 (1997).
Ono, K. & Han, J. The p38 signal transduction pathway: activation and function. Cell. Signal. 12, 1–13 (2000).
Tanaka, N. et al. Differential involvement of p38 mitogen-activated protein kinase kinases MKK3 and MKK6 in T-cell apoptosis. EMBO Rep. 3, 785–791 (2002).
Zhang, J. et al. p38 mitogen-activated protein kinase mediates signal integration of TCR/CD28 costimulation in primary murine T cells. J. Immunol. 162, 3819–3829 (1999).
Rincon, M. et al. Interferon-γ expression by Th1 effector T cells mediated by the p38 MAP kinase signaling pathway. EMBO J. 17, 2817–2829 (1998).
Ward, S.G., Parry, R.V., Matthews, J. & O'Neill, L. A p38 MAP kinase inhibitor SB203580 inhibits CD28-dependent T cell proliferation and IL-2 production. Biochem. Soc. Trans. 25, 304S (1997).
Crawley, J.B. et al. T cell proliferation in response to interleukins 2 and 7 requires p38MAP kinase activation. J. Biol. Chem. 272, 15023–15027 (1997).
Matsuda, S., Moriguchi, T., Koyasu, S. & Nishida, E. T lymphocyte activation signals for interleukin-2 production involve activation of MKK6-p38 and MKK7-SAPK/JNK signaling pathways sensitive to cyclosporin A. J. Biol. Chem. 273, 12378–12382 (1998).
Haeryfar, S.M. & Hoskin, D.W. Selective pharmacological inhibitors reveal differences between Thy-1- and T cell receptor-mediated signal transduction in mouse T lymphocytes. Int. Immunopharmacol. 1, 689–698 (2001).
Jiang, Y. et al. Structure-function studies of p38 mitogen-activated protein kinase. Loop 12 influences substrate specificity and autophosphorylation, but not upstream kinase selection. J. Biol. Chem. 272, 11096–11102 (1997).
Finco, T.S., Kadlecek, T., Zhang, W., Samelson, L.E. & Weiss, A. LAT is required for TCR-mediated activation of PLCγ1 and the Ras pathway. Immunity 9, 617–626 (1998).
Chan, A.C., Desai, D.M. & Weiss, A. The role of protein tyrosine kinases and protein tyrosine phosphatases in T cell antigen receptor signal transduction. Annu. Rev. Immunol. 12, 555–592 (1994).
Williams, B.L. et al. Genetic evidence for differential coupling of Syk family kinases to the T-cell receptor: reconstitution studies in a ZAP-70-deficient Jurkat T-cell line. Mol. Cell. Biol. 18, 1388–1399 (1998).
Steinberg, M. et al. T-cell receptor-induced phosphorylation of the ζ chain is efficiently promoted by ZAP-70 but not Syk. Blood 104, 760–767 (2004).
Salojin, K.V., Zhang, J. & Delovitch, T.L. TCR and CD28 are coupled via ZAP-70 to the activation of the Vav/Rac-1-/PAK-1/p38 MAPK signaling pathway. J. Immunol. 163, 844–853 (1999).
Yu, H., Leitenberg, D., Li, B. & Flavell, R.A. Deficiency of small GTPase Rac2 affects T cell activation. J. Exp. Med. 194, 915–926 (2001).
Salvador, J.M., Mittelstadt, P.R., Belova, G.I., Fornace, A.J., Jr & Ashwell, J.D. The autoimmune suppressor GADD45α inhibits the T cell alternative p38 activation pathway. Nat. Immunol. (27 February 2005) 10.1038/ni1176.
Chi, H., Lu, B., Takekawa, M., Davis, R.J. & Flavell, R.A. GADD45β/GADD45γ and MEKK4 comprise a genetic pathway mediating STAT4-independent IFNγ production in T cells. EMBO J. 23, 1576–1586 (2004).
Wilson, K.P. et al. Crystal structure of p38 mitogen-activated protein kinase. J. Biol. Chem. 271, 27696–27700 (1996).
Weiss, A. & Littman, D.R. Signal transduction by lymphocyte antigen receptors. Cell 76, 263–274 (1994).
Bell, M. & Engelberg, D. Phosphorylation of Tyr-176 of the yeast MAPK Hog1/p38 is not vital for Hog1 biological activity. J. Biol. Chem. 278, 14603–14606 (2003).
Ge, B. et al. MAPKK-independent activation of p38α mediated by TAB1-dependent autophosphorylation of p38α. Science 295, 1291–1294 (2002).
D'Oro, U., Sakaguchi, K., Appella, E. & Ashwell, J.D. Mutational analysis of Lck in CD45-negative T cells: dominant role of tyrosine 394 phosphorylation in kinase activity. Mol. Cell. Biol. 16, 4996–5003 (1996).
Salvador, J.M. et al. Mice lacking the p53-effector gene Gadd45a develop a lupus-like syndrome. Immunity 16, 499–508 (2002).
We thank S. Gutkind for critical review of this manuscript.
About this article
Scientific Reports (2016)