Abstract
The 'classical' NF-kappaB activation pathway proceeds via IkappaB kinase (IKK)-beta/gamma-mediated phosphorylation, induced ubiquitination and the degradation of small IkappaBs. An alternative, NF-kappaB-inducing kinase and IKK-alpha-dependent pathway, which stimulates the processing of NF-kappaB2/p100, has recently been suggested. However, no physiological stimulus has been shown to trigger the activation of this pathway. Here we demonstrate that persistent stimulation with lymphotoxin beta (LT-beta) receptor agonists or lipopolysaccharide (LPS), but not with interleukin-1beta, tumour necrosis factor-alpha or 12-O-tetradecanoylphorbol-13-acetate, induces the generation of p52 DNA-binding complexes by activating the processing of the p100 precursor. Induction of p52 DNA-binding activity is delayed in comparison with p50/p65 complexes and depends on de novo protein synthesis. p100 is constitutively and inducibly polyubiquitinated, and both ubiquitination and p52 generation are coupled to continuing p100 translation. Thus, both LT-beta receptor agonists and LPS induce NF-kappaB/p100 processing to p52 at the level of the ribosome.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing
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Adenocarcinoma / pathology
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B-Lymphocytes / drug effects
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B-Lymphocytes / metabolism
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Breast Neoplasms / pathology
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DNA / metabolism
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Dendritic Cells / drug effects
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Dendritic Cells / metabolism
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Enzyme Activation
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Female
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HeLa Cells / drug effects
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HeLa Cells / metabolism
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Humans
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I-kappa B Proteins / chemistry
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I-kappa B Proteins / genetics
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I-kappa B Proteins / physiology
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Interleukin-1 / pharmacology
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Lipopolysaccharides / pharmacology*
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Lymphotoxin beta Receptor
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Lymphotoxin-alpha / agonists
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Lymphotoxin-alpha / pharmacology*
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Lymphotoxin-beta
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Membrane Proteins / agonists
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Membrane Proteins / pharmacology*
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NF-KappaB Inhibitor alpha
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NF-kappa B / metabolism*
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NF-kappa B p52 Subunit
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Protein Binding / drug effects
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Protein Precursors / metabolism*
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Protein Processing, Post-Translational
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Receptors, Tumor Necrosis Factor / agonists*
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Receptors, Tumor Necrosis Factor / physiology
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / physiology
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Ribosomes / metabolism
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Sequence Deletion
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Tetradecanoylphorbol Acetate / pharmacology
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Trans-Activators / metabolism*
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Transcription Factors
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Tumor Cells, Cultured / drug effects
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Tumor Cells, Cultured / metabolism
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Tumor Necrosis Factor Ligand Superfamily Member 14
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Tumor Necrosis Factor-alpha / pharmacology
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Ubiquitin / metabolism
Substances
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Adaptor Proteins, Signal Transducing
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I-kappa B Proteins
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Interleukin-1
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LTB protein, human
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LTBR protein, human
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Lipopolysaccharides
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Lymphotoxin beta Receptor
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Lymphotoxin-alpha
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Lymphotoxin-beta
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Membrane Proteins
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NF-kappa B
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NF-kappa B p52 Subunit
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NFKBIA protein, human
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PSIP1 protein, human
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Protein Precursors
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Receptors, Tumor Necrosis Factor
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Recombinant Fusion Proteins
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TNFSF14 protein, human
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Trans-Activators
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Transcription Factors
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Tumor Necrosis Factor Ligand Superfamily Member 14
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Tumor Necrosis Factor-alpha
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Ubiquitin
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NF-KappaB Inhibitor alpha
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DNA
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Tetradecanoylphorbol Acetate