The Rel family member P50 mediates cytokine-induced C-reactive protein expression by a novel mechanism.
Transcription of C-reactive protein (CRP) in Hep 3B cells is induced by IL-6, acting through C/EBP isoforms and STAT3. IL-1beta, which alone has no effect, greatly enhances IL-6-induced transcription by unknown mechanisms. Because IL-1beta activates the NF-kappaB system, we explored the effects of overexpressed Rel family members on CRP expression. Unexpectedly, transactivation assays in transiently transfected Hep 3B cells showed p50 overexpression to markedly induce CRP transcription, acting in a region 3' to -86. In the presence of overexpressed p50, IL-1beta induced a 3-fold increase in CRP expression, and responses to IL-6 and to IL-6 plus IL-1beta were 4-fold greater than seen in cells without p50 overexpression. In contrast, overexpressed p65 abolished CRP induction by p50 and by cytokines. EMSA studies demonstrated that recombinant p50 bound to a nonconsensus kappaB site overlapping the proximal C/EBP binding site on the CRP promoter. Mutation of a polypyrimidine tract in the p50-binding site inhibited the transactivating effect of cytokines. P50- but not p65-containing dimers were found in nuclei of Hep 3B cells 18 h after stimulation with IL-1beta, when C/EBPbeta is greatly activated, in the presence or absence of IL-6. These findings suggest that IL-1beta induces nuclear translocation of p50-containing dimers and that p50 interacts with C/EBPbeta activated by both IL-6 and IL-1beta to induce CRP expression.
Cha-Molstad
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H
Department of Biochemistry, Case Western Reserve University, Cleveland, OH 44106, USA.
Agrawal
A
A
Zhang
D
D
Samols
D
D
Kushner
I
I
D020413:3' Untranslated Regions / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D001483:Base Sequence; D002097:C-Reactive Protein / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D002467:Cell Nucleus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016384:Consensus Sequence; D016207:Cytokines / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016328:NF-kappa B / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D052002:NF-kappa B p50 Subunit; D011401:Promoter Regions, Genetic / Q000276:immunology; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D020569:Proto-Oncogene Proteins c-rel / Q000502:physiology*; D020218:Response Elements / Q000276:immunology; D051996:Transcription Factor RelA; D015533:Transcriptional Activation / Q000276:immunology; D014162:Transfection; D014407:Tumor Cells, Cultured
D020413:3' Untranslated Regions; D016207:Cytokines; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D020569:Proto-Oncogene Proteins c-rel; D051996:Transcription Factor RelA; D002097:C-Reactive Protein