Identification of essential regions in the cytoplasmic tail of interleukin-1 receptor accessory protein critical for interleukin-1 signaling.
Interleukin (IL)-1 plays an important role in inflammation and regulation of immune responses. The activated IL-1 receptor complex, which consists of the IL-1 receptor type I and the IL-1 receptor accessory protein (IL-1RAcP), generates multiple cellular responses including NF-kappaB activation, IL-2 secretion, and IL-2 promoter activation. Reconstitution experiments in EL4D6/76 ... cells lacking IL-1RAcP expression and IL-1 responsiveness were used to analyze structure-function relationships of the IL-1RAcP cytoplasmic tail. Mutating a potential tyrosine kinase phosphorylation motif and various conserved amino acid (aa) residues had no effect on IL-1 responsiveness. Truncation analyses revealed that box 3 of the TIR domain was required for NF-kappaB activation, IL-2 production, and c-Jun N-terminal kinase (JNK) activation, whereas IL-2 promoter activation was only partially inhibited. Surprisingly, deletion of aa 527-534 resulted in almost complete loss of all IL-1 responsiveness. Replacement of these aa with alanyl residues did not reconstitute NF-kappaB activation, IL-2 production, or JNK activation but partly restored IL-2 promoter activation. Immunoprecipitation data revealed a strong correlation between MyD88 binding with NF-kappaB activation and IL-2 production but not with IL-2 promoter activation. Taken together, our data indicate that box 3 of IL-1RAcP is critical for IL-1-dependent NF-kappaB activation and stabilization of IL-2 mRNA via JNK, whereas aa 527-534 largely contribute to IL-2 promoter activation.
Mesh Terms:
Amino Acid Sequence, Animals, Blotting, Western, Cell Line, Cytoplasm, Enzyme Activation, Enzyme-Linked Immunosorbent Assay, Gene Deletion, Genes, Reporter, Humans, Interleukin-1, Interleukin-1 Receptor Accessory Protein, Interleukin-2, JNK Mitogen-Activated Protein Kinases, Luciferases, Mice, Mitogen-Activated Protein Kinases, Models, Biological, Molecular Sequence Data, Mutation, NF-kappa B, Plasmids, Precipitin Tests, Promoter Regions, Genetic, Protein Binding, Protein Structure, Tertiary, Proteins, Signal Transduction, Structure-Activity Relationship, Time Factors, Transfection
Amino Acid Sequence, Animals, Blotting, Western, Cell Line, Cytoplasm, Enzyme Activation, Enzyme-Linked Immunosorbent Assay, Gene Deletion, Genes, Reporter, Humans, Interleukin-1, Interleukin-1 Receptor Accessory Protein, Interleukin-2, JNK Mitogen-Activated Protein Kinases, Luciferases, Mice, Mitogen-Activated Protein Kinases, Models, Biological, Molecular Sequence Data, Mutation, NF-kappa B, Plasmids, Precipitin Tests, Promoter Regions, Genetic, Protein Binding, Protein Structure, Tertiary, Proteins, Signal Transduction, Structure-Activity Relationship, Time Factors, Transfection
J. Biol. Chem.
Date: May. 10, 2002
PubMed ID: 11880380
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