RRC ID 12208
Author Miyazawa K, Mori A, Yamamoto K, Okudaira H.
Title Transcriptional roles of CCAAT/enhancer binding protein-beta, nuclear factor-kappaB, and C-promoter binding factor 1 in interleukin (IL)-1beta-induced IL-6 synthesis by human rheumatoid fibroblast-like synoviocytes.
Journal J Biol Chem
Abstract The involvement of interleukin (IL)-6 in the pathogenesis of rheumatoid arthritis (RA) has been recently demonstrated. IL-1beta stimulated rheumatoid fibroblast-like synoviocytes (FLSs) to produce IL-6 in a concentration- and time-dependent manner. In the present study we investigated how the IL-6 promoter is transcriptionally regulated in rheumatoid FLSs in response to a physiologically relevant mediator of inflammation, IL-1beta. Deletion analysis showed that the IL-6 promoter is regulated by two positive elements (located at -159 to -142 base pairs (bp) and -77 to -59 bp). Electrophoretic mobility shift assays revealed that CCAAT/enhancer binding protein-beta (C/EBPbeta) binding to nucleotides -159 to -142 bp was constitutively present. The probe corresponding to nucleotides -77 to -59 bp gave three positive bands. The two slower migrating bands were induced by IL-1beta and comprised an nuclear factor (NF)-kappaB p50/p65 heterodimer and a p65/p65 homodimer. The faster migrating band was constitutively expressed and identified as Epstein-Barr virus C-promoter binding factor 1, CBF1. Site-specific mutagenesis analysis demonstrated that the NF-kappaB and CBF1 binding elements regulated inducible activity of the IL-6 promoter in response to IL-1beta stimulation, whereas the C/EBPbeta binding element mainly regulated basal activity. We also provide the first evidence that CBF1 functions as a positive regulator of human IL-6 gene transcription.
Volume 273(13)
Pages 7620-7
Published 1998-3-27
DOI 10.1074/jbc.273.13.7620
PII S0021-9258(18)61738-7
PMID 9516466
MeSH Arthritis, Rheumatoid / metabolism* CCAAT-Enhancer-Binding Proteins DNA-Binding Proteins / metabolism* Fibroblasts / metabolism Gene Expression Regulation / drug effects Humans Immunoglobulin J Recombination Signal Sequence-Binding Protein Interleukin-1 / pharmacology* Interleukin-6 / biosynthesis* Interleukin-6 / genetics NF-kappa B / metabolism* Nuclear Proteins / metabolism* Promoter Regions, Genetic Recombinant Proteins / metabolism Synovial Membrane / cytology Synovial Membrane / drug effects Synovial Membrane / metabolism* Transcription Factors / metabolism* Transcription, Genetic
IF 4.238
Times Cited 68
DNA material pGEMhIL-6 GT (RDB01211)