IL-1beta induces eotaxin gene transcription in A549 airway epithelial cells through NF-kappaB

Eotaxin is an asthma-related C-C chemokine that is produced in response to interleukin-1beta (IL-1beta). We detected an increase in newly transcribed eotaxin mRNA in IL-1beta-stimulated airway epithelial cells. Transient transfection assays using promoter-reporter constructs identified a region as e...

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Bibliographic Details
Published in:American journal of physiology. Lung cellular and molecular physiology 2000-12, Vol.279 (6), p.L1058
Main Authors: Jedrzkiewicz, S, Nakamura, H, Silverman, E S, Luster, A D, Mansharamani, N, In, K H, Tamura, G, Lilly, C M
Format: Article
Language:English
Subjects:
Adenocarcinoma, Bronchiolo-Alveolar
Chemokine CCL11
Chemokines, CC
Cytokines - genetics
Epithelial Cells - drug effects
Epithelial Cells - physiology
Gene Expression Regulation - drug effects
Genes, Reporter
Genetic Complementation Test
Humans
Interleukin-1 - pharmacology
Lung Neoplasms
Mutagenesis, Site-Directed - physiology
NF-kappa B - genetics
NF-kappa B - metabolism
Promoter Regions, Genetic - physiology
Respiratory Mucosa - cytology
Respiratory Mucosa - physiology
RNA, Messenger - metabolism
Transcriptional Activation - drug effects
Tumor Cells, Cultured
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Summary:Eotaxin is an asthma-related C-C chemokine that is produced in response to interleukin-1beta (IL-1beta). We detected an increase in newly transcribed eotaxin mRNA in IL-1beta-stimulated airway epithelial cells. Transient transfection assays using promoter-reporter constructs identified a region as essential for IL-1beta-induced increases in eotaxin transcription. Using site-directed mutagenesis, we found that a nuclear factor-kappaB (NF-kappaB) site located 46 bp upstream from the transcriptional start site was both necessary and sufficient for IL-1beta induction of reporter construct activity. Electrophoretic mobility shift assay demonstrated that IL-1beta-stimulated airway epithelial cells produced p50 and p65 protein that bound this site in a sequence-specific manner. The functional importance of the NF-kappaB site was demonstrated by coexpression experiments in which increasing doses of p65 expression vector were directly associated with reporter activity exclusively in constructs with an intact NF-kappaB site (r(2) = 0.97, P = 0.002). Moreover, IL-1beta-induced increases in eotaxin mRNA expression are inhibited by inhibitors of NF-kappaB. Our findings implicate NF-kappaB and its binding sequence in IL-1beta-induced transcriptional activation of the eotaxin gene.
ISSN:1040-0605