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CARMA1 is a critical lipid raft–associated regulator of TCR-induced NF-κB activation

Abstract

CARMA1 is a lymphocyte-specific member of the membrane-associated guanylate kinase (MAGUK) family of scaffolding proteins, which coordinate signaling pathways emanating from the plasma membrane. CARMA1 interacts with Bcl10 via its caspase-recruitment domain (CARD). Here we investigated the role of CARMA1 in T cell activation and found that T cell receptor (TCR) stimulation induced a physical association of CARMA1 with the TCR and Bcl10. We found that CARMA1 was constitutively associated with lipid rafts, whereas cytoplasmic Bcl10 translocated into lipid rafts upon TCR engagement. A CARMA1 mutant, defective for Bcl10 binding, had a dominant-negative (DN) effect on TCR-induced NF-κB activation and IL-2 production and on the c-Jun NH2-terminal kinase (Jnk) pathway when the TCR was coengaged with CD28. Together, our data show that CARMA1 is a critical lipid raft–associated regulator of TCR-induced NF-κB activation and CD28 costimulation–dependent Jnk activation.

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Figure 1: Association of CARMA1 with the TCR complex and with Bcl10 after T cell activation.
Figure 2: Colocalization of CARMA1 and Bcl10 with the TCR after CD3-aggregation.
Figure 3: Differential lipid raft localization of CARMA1 and Bcl10 in resting and CD3-triggered T cells.
Figure 4: Inhibitory effect of CARD-mutated CARMA1 on TCR-mediated NF-κB activation and IL-2 production.
Figure 5: Effects of DN CARMA1 on TCR-triggered signaling pathways.
Figure 6: Effects of DN CARMA1 on PMA + ionomycin and CD28 costimulation–induced signaling pathways.

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Acknowledgements

Supported by grants from the Swiss Cancer League (to M. T. and J. T.) and by the Swiss Academy of Medical Sciences (to O. G). C. B. and D. L. were supported by grants from the Swiss National Science Foundation and the Giorgi Cavaglieri Foundation; S. V. was supported by grants from La Ligue contre le Cancer, La Fondation pour la Recherche Médicale and the Giorgio Cavaglieri Foundation. We thank S. Levrand for technical assistance, P. Zaech for help with FACS analysis and K. Burns, F. Martinon and M. Thurau for critical review of the manuscript.

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Correspondence to Margot Thome.

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Gaide, O., Favier, B., Legler, D. et al. CARMA1 is a critical lipid raft–associated regulator of TCR-induced NF-κB activation. Nat Immunol 3, 836–843 (2002). https://doi.org/10.1038/ni830

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