Phosphorylation of CARMA1 by HPK1 is critical for NF-kappaB activation in T cells.
Activation of the NF-kappaB pathway in T cells is required for induction of an adaptive immune response. Hematopoietic progenitor kinase (HPK1) is an important proximal mediator of T-cell receptor (TCR)-induced NF-kappaB activation. Knock-down of HPK1 abrogates TCR-induced IKKbeta and NF-kappaB activation, whereas active HPK1 leads to increased IKKbeta activity in T cells. Yet, the precise molecular mechanism of this process remains elusive. Here, we show that HPK1-mediated NF-kappaB activation is dependent on the adaptor protein CARMA1. HPK1 interacts with CARMA1 in a TCR stimulation-dependent manner and phosphorylates the linker region of CARMA1. Interestingly, the putative HPK1 phosphorylation sites in CARMA1 are different from known PKC consensus sites. Mutations of residues S549, S551, and S552 in CARMA1 abrogated phosphorylation of a CARMA1-linker construct by HPK1 in vitro. In addition, CARMA1 S551A or S5549A/S551A point mutants failed to restore HPK1-mediated and TCR-mediated NF-kappaB activation and IL-2 expression in CARMA1-deficient T cells. Thus, we identify HPK1 as a kinase specific for CARMA1 and suggest HPK1-mediated phosphorylation of CARMA1 as an additional regulatory mechanism tuning the NF-kappaB response upon TCR stimulation.
Brenner
Dirk
D
Division of Immunogenetics, Tumor Immunology Program, German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany.
Brechmann
Markus
M
Röhling
Simone
S
Tapernoux
Myriam
M
Mock
Thomas
T
Winter
Dominic
D
Lehmann
Wolf D
WD
Kiefer
Friedemann
F
Thome
Margot
M
Krammer
Peter H
PH
Arnold
Rüdiger
R
D000818:Animals; D001665:Binding Sites; D053468:CARD Signaling Adaptor Proteins / Q000235:genetics / Q000378:metabolism*; D019556:COS Cells; D002460:Cell Line; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D006162:Guanylate Cyclase / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D019169:Jurkat Cells; D009154:Mutation; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000378:metabolism*; D014162:Transfection
D053468:CARD Signaling Adaptor Proteins; D016328:NF-kappa B; D034741:RNA, Small Interfering; D011948:Receptors, Antigen, T-Cell; C102102:hematopoietic progenitor kinase 1; D017346:Protein Serine-Threonine Kinases; C426242:CARD11 protein, human; D006162:Guanylate Cyclase
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