Abstract
Transforming growth factor-beta-activated kinase 1 (TAK1) plays an essential role in the tumor necrosis factor alpha (TNFalpha)- and interleukin-1beta (IL-1beta)-induced IkappaB kinase (IKK)/nuclear factor-kappaB (NF-kappaB) and c-Jun N-terminal kinase (JNK)/activator protein 1 (AP-1) activation. Here we report that TNFalpha and IL-1beta induce Lys(63)-linked TAK1 polyubiquitination at the Lys(158) residue within the kinase domain. Tumor necrosis factor receptor-associated factors 2 and 6 (TRAF2 and -6) act as the ubiquitin E3 ligases to mediate Lys(63)-linked TAK1 polyubiquitination at the Lys(158) residue in vivo and in vitro. Lys(63)-linked TAK1 polyubiquitination at the Lys(158) residue is required for TAK1-mediated IKK complex recruitment. Reconstitution of TAK1-deficient mouse embryo fibroblast cells with TAK1 wild type or a TAK1 mutant containing a K158R mutation revealed the importance of this site in TNFalpha and IL-1beta-mediated IKK/NF-kappaB and JNK/AP-1 activation as well as IL-6 gene expression. Our findings demonstrate that Lys(63)-linked polyubiquitination of TAK1 at Lys(158) is essential for its own kinase activation and its ability to mediate its downstream signal transduction pathways in response to TNFalpha and IL-1beta stimulation.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Substitution
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Animals
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Cell Line
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Embryo, Mammalian / metabolism
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Fibroblasts / metabolism
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Humans
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I-kappa B Kinase / genetics
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I-kappa B Kinase / metabolism*
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Interleukin-1beta / metabolism*
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Interleukin-1beta / pharmacology
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Interleukin-6 / biosynthesis
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Interleukin-6 / genetics
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JNK Mitogen-Activated Protein Kinases / genetics
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JNK Mitogen-Activated Protein Kinases / metabolism*
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Lysine / genetics
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Lysine / metabolism*
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MAP Kinase Kinase Kinases / genetics
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MAP Kinase Kinase Kinases / metabolism*
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Mice
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Mice, Mutant Strains
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Mutation, Missense
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NF-kappa B / genetics
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NF-kappa B / metabolism*
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Signal Transduction / drug effects
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Signal Transduction / physiology
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TNF Receptor-Associated Factor 2 / genetics
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TNF Receptor-Associated Factor 2 / metabolism
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TNF Receptor-Associated Factor 6 / genetics
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TNF Receptor-Associated Factor 6 / metabolism
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Transcription Factor AP-1 / genetics
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Transcription Factor AP-1 / metabolism*
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Tumor Necrosis Factor-alpha / metabolism*
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Tumor Necrosis Factor-alpha / pharmacology
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Ubiquitin-Protein Ligases / genetics
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Ubiquitin-Protein Ligases / metabolism
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Ubiquitination / drug effects
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Ubiquitination / physiology*
Substances
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IL6 protein, human
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Interleukin-1beta
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Interleukin-6
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NF-kappa B
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TNF Receptor-Associated Factor 2
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TNF Receptor-Associated Factor 6
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Transcription Factor AP-1
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Tumor Necrosis Factor-alpha
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Ubiquitin-Protein Ligases
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I-kappa B Kinase
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JNK Mitogen-Activated Protein Kinases
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MAP Kinase Kinase Kinases
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MAP kinase kinase kinase 7
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Lysine