USP2a positively regulates TCR-induced NF-κB activation by bridging MALT1-TRAF6

Abstract

The paracaspase MALT1 is essential for the activation of NF-κB in response to T cell receptor (TCR) stimulation. It recruits downstream TRAF6 and activates the E3 ligase activity of TRAF6 to polyubiquitinate several targets, which ultimately leads to NF-κB activation. Here we identified ubiquitin-specific protease 2a (USP2a) as a MALT1-associated protein by biochemical affinity purification. Endogenous USP2a constitutively interacted with TRAF6, but dynamically interacted with MALT1 and CARMA1 in a stimulation-dependent manner. RNA interference (RNAi)-mediated silencing of USP2a attenuated TCR-induced NF-κB activation and production of interleukin-2 (IL-2). In addition, the ubiquitination of MALT1 and TRAF6 were both suppressed by USP2a knockdown. By knockdown and reconstitution assays, we found that USP2a mediated the interaction between MALT1 and TRAF6 in a catalytic activity- dependent manner. Furthermore, USP2a deSUMOylated TRAF6. Our findings implicate that USP2a plays an important role in TCR signaling by deSUMOylating TRAF6 and mediating TRAF6-MALT1 interaction.

Keywords

USP2a / MALT1 / TRAF6 / T cell activation / NF-κB

References

[1] Alberola-Ila, J., Takaki, S., Kerner, J.D., and Perlmutter, R.M. (1997). Differential signaling by lymphocyte antigen receptors .Annu Rev Immunol 15, 125-154 .10.1146/annurev.immunol.15.1.125
[2] Bergink, S., and Jentsch, S. (2009). Principles of ubiquitin and SUMO modifications in DNA repair. Nature 458, 461-467 .10.1038/nature07963
[3] Coornaert, B., Baens, M., Heyninck, K., Bekaert, T., Haegman, M., Staal, J., Sun, L., Chen, Z.J., Marynen, P., and Beyaert, R. (2008). T cell antigen receptor stimulation induces MALT1 paracaspase-mediated cleavage of the NF-kappaB inhibitor A20. Nat Immunol 9, 263-271 .10.1038/ni1561
[4] Deng, L., Wang, C., Spencer, E., Yang, L., Braun, A., You, J., Slaughter, C., Pickart, C., and Chen, Z.J. (2000). Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain. Cell 103, 351-361 .10.1016/S0092-8674(00)00126-4
[5] Gaide, O., Favier, B., Legler, D.F., Bonnet, D., Brissoni, B., Valitutti, S., Bron, C., Tschopp, J., and Thome, M. (2002). CARMA1 is a critical lipid raft-associated regulator of TCR-induced NF-kappa B activation. Nat Immunol 3, 836-843 .10.1038/ni830
[6] Ghosh, S., May, M.J., and Kopp, E.B. (1998). NF-kappa B and Rel proteins: evolutionarily conserved mediators of immune responses. Annu Rev Immunol 16, 225-260 .10.1146/annurev.immunol.16.1.225
[7] Hannoun, Z., Greenhough, S., Jaffray, E., Hay, R.T., and Hay, D.C. (2010). Post-translational modification by SUMO. Toxicology 278, 288-293 .10.1016/j.tox.2010.07.013
[8] He, X., Li, Y., Li, C., Liu, L.J., Zhang, X.D., Liu, Y., and Shu, H.B. (2012). USP2a negatively regulates IL-1beta- and virus-induced NF-kappa B activation by deubiquitinating TRAF6. J Mol Cell Biol. (In Press).10.1093/jmcb/mjs024
[9] Li, Y., Chen, R., Zhou, Q., Xu, Z., Li, C., Wang, S., Mao, A., Zhang, X., He, W., and Shu, H.B. (2012). LSm14A is a processing body-associated sensor of viral nucleic acids that initiates cellular antiviral response in the early phase of viral infection. Proc Natl Acad Sci U S A 109, 11770-11775 .10.1073/pnas.1203405109
[10] Liu, Y., Song, R., Gao, Y., Li, Y., Wang, S., Liu, H.Y., Wang, Y., Hu, Y.H., and Shu, H.B. (2012). Protein kinase C-delta negatively regulates T cell receptor-induced NF-kappaB activation by inhibiting the assembly of CARMA1 signalosome. J Biol Chem 287, 20081-20087 .10.1074/jbc.M111.335463
[11] Mahul-Mellier, A.L., Pazarentzos, E., Datler, C., Iwasawa, R., AbuAli, G., Lin, B., and Grimm, S. (2012). De-ubiquitinating protease USP2a targets RIP1 and TRAF2 to mediate cell death by TNF. Cell Death Differ 19, 891-899 .10.1038/cdd.2011.185
[12] Metzig, M., Nickles, D., Falschlehner, C., Lehmann-Koch, J., Straub, B.K., Roth, W., and Boutros, M. (2011). An RNAi screen identifies USP2 as a factor required for TNF-alpha-induced NF-kappaB signaling. Int J Cancer 129, 607-618 .10.1002/ijc.26124
[13] Oeckinghaus, A., Wegener, E., Welteke, V., Ferch, U., Arslan, S.C., Ruland, J., Scheidereit, C., and Krappmann, D. (2007). Malt1 ubiquitination triggers NF-kappaB signaling upon T-cell activation. EMBO J 26, 4634-4645 .10.1038/sj.emboj.7601897
[14] Oh, K.H., Yang, S.W., Park, J.M., Seol, J.H., Iemura, S., Natsume, T., Murata, S., Tanaka, K., Jeon, Y.J., and Chung, C.H. (2011). Control of AIF-mediated cell death by antagonistic functions of CHIP ubiquitin E3 ligase and USP2 deubiquitinating enzyme. Cell Death Differ 18, 1326-1336 .10.1038/cdd.2011.3
[15] Pham, L.V., Zhou, H.J., Lin-Lee, Y.C., Tamayo, A.T., Yoshimura, L.C., Fu, L., Darnay, B.G., and Ford, R.J. (2008). Nuclear tumor necrosis factor receptor-associated factor 6 in lymphoid cells negatively regulates c-Myb-mediated transactivation through small ubiquitinrelated modifier-1 modification. J Biol Chem 283, 5081-5089 .10.1074/jbc.M706307200
[16] Pomerantz, J.L., Denny, E.M., and Baltimore, D. (2002). CARD11 mediates factor-specific activation of NF-kappaB by the T cell receptor complex. EMBO J 21, 5184-5194 .10.1093/emboj/cdf505
[17] Rebeaud, F., Hailfinger, S., Posevitz-Fejfar, A., Tapernoux, M., Moser, R., Rueda, D., Gaide, O., Guzzardi, M., Iancu, E.M., Rufer, N., . (2008). The proteolytic activity of the paracaspase MALT1 is key in T cell activation. Nat Immunol 9, 272-281 .10.1038/ni1568
[18] Ruefli-Brasse, A.A., French, D.M., and Dixit, V.M. (2003). Regulation of NF-kappaB-dependent lymphocyte activation and development by paracaspase. Science 302, 1581-1584 .10.1126/science.1090769
[19] Ruland, J., Duncan, G.S., Elia, A., del Barco Barrantes, I., Nguyen, L., Plyte, S., Millar, D.G., Bouchard, D., Wakeham, A., Ohashi, P.S., . (2001). Bcl10 is a positive regulator of antigen receptor-induced activation of NF-kappaB and neural tube closure. Cell 104, 33-42 .10.1016/S0092-8674(01)00189-1
[20] Shan, J., Zhao, W., and Gu, W. (2009). Suppression of cancer cell growth by promoting cyclin D1 degradation. Mol Cell 36, 469-476 .10.1016/j.molcel.2009.10.018
[21] Shin, E.J., Shin, H.M., Nam, E., Kim, W.S., Kim, J.H., Oh, B.H., and Yun, Y. (2012). DeSUMOylating isopeptidase: a second class of SUMO protease. EMBO Rep 13, 339-346 .10.1038/embor.2012.3
[22] Stevenson, L.F., Sparks, A., Allende-Vega, N., Xirodimas, D.P., Lane, D.P., and Saville, M.K. (2007). The deubiquitinating enzyme USP2a regulates the p53 pathway by targeting Mdm2. EMBO J 26, 976-986 .10.1038/sj.emboj.7601567
[23] Sun, L., and Chen, Z.J. (2004). The novel functions of ubiquitination in signaling. Curr Opin Cell Biol 16, 119-126 .10.1016/j.ceb.2004.02.005
[24] Sun, L., Deng, L., Ea, C.K., Xia, Z.P., and Chen, Z.J. (2004). The TRAF6 ubiquitin ligase and TAK1 kinase mediate IKK activation by BCL10 and MALT1 in T lymphocytes. Mol Cell 14, 289-301 .10.1016/S1097-2765(04)00236-9
[25] Thome, M., and Weil, R. (2007). Post-translational modifications regulate distinct functions of CARMA1 and BCL10. Trends Immunol 28, 281-288 .10.1016/j.it.2007.04.004
[26] Wang, D., You, Y., Case, S.M., McAllister-Lucas, L.M., Wang, L., DiStefano, P.S., Nunez, G., Bertin, J., and Lin, X. (2002). A requirement for CARMA1 in TCR-induced NF-kappa B activation. Nat Immunol 3, 830-835 .10.1038/ni824
[27] Wegener, E., Oeckinghaus, A., Papadopoulou, N., Lavitas, L., Schmidt-Supprian, M., Ferch, U., Mak, T.W., Ruland, J., Heissmeyer, V., and Krappmann, D. (2006). Essential role for IkappaB kinase beta in remodeling Carma1-Bcl10-Malt1 complexes upon T cell activation. Mol Cell 23, 13-23 .10.1016/j.molcel.2006.05.027