title: TNFR-associated factor 6 regulates TCR signaling via interaction with and modification of LAT adapter.
abstract: TNFR-associated factor (TRAF)6 is an essential ubiquitin E3 ligase in immune responses, but its function in adaptive immunity is not well understood. In this study, we show that TRAF6 is recruited to the peripheral ring of the T cell immunological synapse in Jurkat T cells or human primary CD4(+) T cells conjugated with staphylococcal enterotoxin E-pulsed B cells. This recruitment depends on TRAF6 interacting with linker for activation of T cells (LAT) via its TRAF domain. Although LAT was indispensable for TCR/CD28-induced TRAF6 ubiquitination and its ligase activity, RNA interference-induced TRAF6 knockdown in T cells decreased TCR/CD28-induced LAT ubiquitination, tyrosine phosphorylation, and association with tyrosine kinase ZAP70. Overexpression of TRAF6 or its catalytically inactive form C70A promoted and decreased, respectively, LAT tyrosine phosphorylation upon stimulation. Moreover, LAT was ubiquitinated at Lys(88) by TRAF6 via K63-linked chain. In addition, TRAF6 was required for and synergized with LAT to promote the TCR/CD28-induced activation of NFAT. These results reveal a novel function and mechanism of TRAF6 action in the TCR-LAT signaling pathway distinct from its role in TCR-induced NF-κB activation, indicating that LAT also plays an adapter role in TCR/CD28-induced activation of TRAF6.
authors.affiliation: State Key Laboratory of Biocontrol, Key Laboratory of Gene Engineering of Ministry of Education, School of Life Sciences, Sun Yat-sen University, Guangzhou 510006, China.
authors.forename: Ji-Ji
authors.initials: JJ
authors.lastname: Xie
authors.forename: Jia-Qi
authors.initials: JQ
authors.lastname: Liang
authors.forename: Liang-Hui
authors.initials: LH
authors.lastname: Diao
authors.forename: Amnon
authors.initials: A
authors.lastname: Altman
authors.forename: Yingqiu
authors.initials: Y
authors.lastname: Li
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D018106:CD28 Antigens / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D019169:Jurkat Cells; D008565:Membrane Proteins / Q000378:metabolism*; D010766:Phosphorylation / Q000276:immunology; D061251:Primary Cell Culture; D025941:Protein Interaction Mapping / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000502:physiology*; D015398:Signal Transduction / Q000276:immunology*; D048029:TNF Receptor-Associated Factor 6 / Q000172:deficiency / Q000378:metabolism / Q000502:physiology*; D054875:Ubiquitination / Q000276:immunology
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018106:CD28 Antigens; C110810:LAT protein, human; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; D048029:TNF Receptor-Associated Factor 6