Abstract
The Ras signaling pathway plays a critical role in thymopoiesis and T cell activation, but the mechanism of Ras regulation is controversial. At least one mode of Ras regulation in T cells involves the messenger diacylglycerol (DAG). RasGRP, a Ras activator with a DAG-binding C1 domain, is expressed in T cells and thymocytes. Here we show that thymi of RasGRP-null mutant mice have approximately normal numbers of immature thymocytes but a marked deficiency of mature, single-positive (CD4+CD8− and CD4−CD8+) thymocytes. In Ras signaling and proliferation assays, mutant thymocytes showed a complete lack of response to DAG analogs or T cell receptor (TCR) stimulation by antibodies. Thus, TCR and DAG are linked through RasGRP to Ras signaling.
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References
Crabtree, G. R. Contingent genetic regulatory events in T lymphocyte activation. Science 243, 355–361 (1989).
Alberola-Ila, J., Takaki, S., Kerner, J. D. & Perlmutter, R. M. Differential signaling by lymphocyte antigen receptors. Annu. Rev. Immunol. 15, 125–154 (1997).
Downward, J., Graves, J. D., Warne, P. H., Rayter, S. & Cantrell, D. A. Stimulation of p21ras upon T-cell activation. Nature 346, 719– 723 (1990).
Egan, S. E. et al. Association of Sos Ras exchange protein with Grb2 is implicated in tyrosine kinase signal transduction and transformation. Nature 363, 45–51 (1993).
Ebinu, J. O. et al. RasGRP, a Ras guanyl nucleotide releasing protein with calcium and diacylglycerol-binding motifs. Science 280, 1082–1086 (1998).
Kawasaki, H. et al. A Rap guanine nucleotide exchange-factor enriched highly in the basal ganglia. Proc. Natl Acad. Sci. USA 95, 13278–13283 (1998).
Tognon, C. E. et al. Regulation of RasGRP via a phorbol ester-responsive C1 domain . Mol. Cell. Biol. 18, 6995– 7008 (1998).
Yamashita, S. et al. CalDAG-GEFIII activation of Ras, R-Ras, and Rap1. J. Biol. Chem. 275, 25488–25493 (2000).
Ebinu, J. O. et al. RasGRP links T-cell receptor signaling to Ras. Blood 95, 3199–3203 (2000).
von Boehmer, H. & Fehling, H. J. Structure and function of the pre-T cell receptor. Ann. Rev. Immunol. 15, 433–452 (1997).
Mombaerts, P. et al. RAG-1-deficient mice have no mature B and T lymphocytes. Cell 68, 869–877 (1992).
Molina, T. J. et al. Profound block in thymocyte development in mice lacking p56lck . Nature 357, 161–164 (1992).
Negishi, I. et al. Essential role for ZAP-70 in both positive and negative selection of thymocytes. Nature 376, 435– 438 (1995).
Zhang, W. et al. Essential role of Lat in T cell development. Immunity 10, 323–332 (1999).
Clements, J. L. et al. Requirement for the leukocyte-specific adapter protein SLP-76 for normal T cell development. Science 281, 416–419 (1998).
Swan, K. A. et al. Involvement of p21ras distinguishes positive and negative selection in thymocytes. EMBO J. 14, 276 –285 (1995).
O'Shea, C. C., Crompton, T., Rosewell, I. R., Hayday, A. C. & Owen, M. J. Raf regulates positive selection . Eur. J. Immunol. 26, 2350– 2355 (1996).
Crompton, T., Gilmour, K. C. & Owen, M. J. The MAP kinase pathway controls differentiation from double-negative to double-positive thymocyte. Cell 86, 243–251 (1996).
Sharp, L. L., Schwarz, D. A., Bott, C. M., Marshall, C. J. & Hedrick, S. M. The influence of the MAPK pathway on T cell lineage commitment. Immunity 7, 609–618 (1997).
Pages, G. et al. Defective thymocyte maturation in p44 MAP kinase (Erk1) knockout mice. Science 286, 1374– 1377 (1999).
Barton, K. et al. Defective thymocyte proliferation and Il-2 production in transgenic mice expressing a dominant-negative form of CREB. Nature 379, 81–85 (1996).
Pettit, G. R., Herald, C. L., Doubek, D. L. & Herald, D. L. Isolation and structure of bryostatin 1. J. Am. Chem. Soc. 104, 6846–6848 (1982).
Lorenzo, P. S., Beheshti, M., Pettit, G. R., Stone, J. C. & Blumberg, P. M. The guanine nucleotide exchange factor RasGRP is a high affinity target for diacylglycerol and phorbol esters . Mol. Pharmacol. 57, 840– 846 (2000).
Hsueh, Y.-P., Liang, H.-E., Ng, S.-Y. & Lai, M.-Z. CD28-costimulation activates cyclic AMP-responsive element binding protein in T lymphocytes. J. Immunol. 158, 85–93 (1997).
Muthusamy, N. & Leiden, J. M. A protein kinase C-, Ras-, and RSK2-dependent signal transduction pathway activates the cAMP-responsive element-binding protein transcription factor following T cell receptor engagement. J. Biol. Chem. 273, 22841–22847 (1998).
Han, J., Lee, J.-D., Bibbs, L. & Ulevitch, R. J. A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. Science 265, 808–811 (1994).
Iwashima, M., Irving, B. A., vanOers, N. S. C., Chan, A. C. & Weiss, A. Sequential interactions of the TCR with two distinct cytoplasmic tyrosine kinases. Science 263, 1136–1139 (1994).
Peterson, E. J., Clements, J. L., Fan, N. & Koretzky, G. A. Adaptor proteins in lymphocyte antigen-receptor signaling. Curr. Opin. Immunol. 10, 337–344 (1998).
Noh, D. Y., Shin, S. H. & Rhee, S. G. Phosphoinositide-specific phospholipase C and mitogenic signaling. Biochim. Biophys. Acta 1242, 99–114 (1995).
Izquierdo, M., Downward, J., Graves, J. D. & Cantrell, D. A. The role of protein kinase C in T cell antigen receptor regulation of p21ras: evidence that two p21ras regulatory pathways coexist in T cells. Mol. Cell. Biol. 12, 3305–3312 (1992).
Buday, L., Egan, S. E., Viciana, P. R., Cantrell, D. A. & Downward, J. A complex of Grb2 adaptor protein, Sos exchange factor, and a 36-kDa membrane-bound tyrosine phosphoprotein is implicated in Ras activation in T cells. J. Biol. Chem. 269, 9019–9023 (1994).
Salojin, K. V., Zhang, J., Meagher, C. & Delovitch, T. L. ZAP-70 is essential for the T cell antigen receptor-induced plasma membrane targeting of SOS and Vav in T cells. J. Biol. Chem. 275, 5966–5975 (1999).
Holsinger, L. J., Spencer, D. M., Austin, D. J., Schreiber, S. L. & Crabtree, G. R. Signal transduction in T lymphocytes using a conditional allele of Sos. Proc. Natl Acad. Sci. USA 92, 9810–9814 (1995).
Cheng, A. M. et al. The Syk and ZAP-70 SH2-containing tyrosine kinases are implicated in pre-T cell receptor signaling. Proc. Natl Acad. Sci. USA 94, 9797–9801 (1997).
Groves, T. et al. Fyn can partially substitute for Lck in T lymphocyte development . Immunity 5, 417–428 (1996).
van Oers, N. S., Lowin-Kropf, B., Finlay, D., Connolly, K. & Weiss, A. Alpha beta T cell development is abolished in mice lacking both Lck and Fyn protein tyrosine kinases. Immunity 5, 429–436 (1996).
Rodewald, H. R., Ogawa, M., Haller, C., Waskow, C. & DiSanto, J. P. Pro-thymocyte expansion by c-kit and the common cytokine receptor γ chain is essential for repertoire formation. Immunity 6, 265–272 (1997).
Robertson, E. J. in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach (ed. Robertson, E. J.) 112–117 (Oxford, IRL, 1987).
Bottorff, D., Ebinu, J. & Stone, J. C. RasGRP, a Ras activator: mouse and human cDNA sequences and chromosomal positions. Mamm. Genome 10, 358–361 (1998).
Acknowledgements
We thank X. H. Xien for help with developing the NX1 cell line, D. Rutkowski for FACs analysis, and P. Blumberg for bryostatin 1. Supported by grants from the Medical Research Council and the National Cancer Institute (Canada) to J. C. S.
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Dower, N., Stang, S., Bottorff, D. et al. RasGRP is essential for mouse thymocyte differentiation and TCR signaling . Nat Immunol 1, 317–321 (2000). https://doi.org/10.1038/79766
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DOI: https://doi.org/10.1038/79766
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