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PMID: 7684503 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Regulation of the interleukin-1 beta (IL-1 beta) gene by mycobacterial components and lipopolysaccharide is mediated by two nuclear factor-IL6 motifs.

Molecular and cellular biology ·Vol. 13 ·No. 6 ·1993-06-00 ·Pages 3831-7

Zhang Y, Rom WN

Abstract

The cytokines interleukin-1 beta (IL-1 beta) and tumor necrosis factor alpha (TNF-alpha) are released by mononuclear phagocytes in vitro after stimulation with mycobacteria and are considered to mediate pathophysiologic events, including granuloma formation and systemic symptoms. We demonstrated that the Mycobacterium tuberculosis cell wall component lipoarabinomannan (LAM) is a very potent inducer of IL-1 beta gene expression in human monocytes and investigated the mechanism of this effect. We localized the LAM-, lipopolysaccharide (LPS)-, and TNF-alpha-inducible promoter activity to a -131/+15 (positions -131 to +15) DNA fragment of the IL-1 beta gene by deletion analysis and chloramphenicol acetyltransferase assay. Within this DNA fragment, there were two novel 9-bp motifs (-90/-82 and -40/-32) with high homology to the nuclear factor-IL6 (NF-IL6) binding site. Site-directed mutagenesis demonstrated that the two NF-IL-6 motifs could be independently activated by LAM, LPS, or TNF-alpha and that they acted in an orientation-independent manner. DNA mobility shift assay revealed specific binding of nuclear protein(s) from LAM-, LPS-, or TNF-alpha-stimulated THP-1 cells to the NF-IL6 motifs. We conclude that the two NF-IL6 sites mediate induction of IL-1 beta in response to the stimuli LAM, LPS, and TNF-alpha.

MeSH Terms
Base Sequence Binding Sites CCAAT-Enhancer-Binding Proteins Cells, Cultured Chloramphenicol O-Acetyltransferase/genetics,metabolism DNA/genetics,metabolism DNA-Binding Proteins/physiology Gene Expression Regulation/drug effects,physiology Genomic Library Humans Interleukin-1/genetics Leukemia Lipopolysaccharides/pharmacology Molecular Sequence Data Monocytes/physiology Mycobacterium tuberculosis Nuclear Proteins/physiology Promoter Regions, Genetic RNA/blood,genetics,isolation & purification Recombinant Proteins/pharmacology Sequence Deletion Transcription Factors/physiology Transfection Tumor Cells, Cultured Tumor Necrosis Factor-alpha/pharmacology
Chemicals
CCAAT-Enhancer-Binding Proteins DNA-Binding Proteins Interleukin-1 Lipopolysaccharides Nuclear Proteins Recombinant Proteins Transcription Factors Tumor Necrosis Factor-alpha lipoarabinomannan RNA DNA Chloramphenicol O-Acetyltransferase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Zhang Y
Department of Medicine, Bellevue Hospital Center, New York University Medical Center, New York 10016.
Rom W N
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41 references, click to expand
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.6.3831-3837.1993
Published
1993-06-00
Pages
3831-7
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359872
Subset
IM
Grants
NCRR NIH HHS · M01 RR00096-2851 · United States
External Links
PubMed source DOI: 10.1128/mcb.13.6.3831-3837.1993 PMC Full Text View journal details
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