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Identification of transcription factors regulating CTNNAL1 expression in human bronchial epithelial cells.
- Source :
-
PloS one [PLoS One] 2012; Vol. 7 (2), pp. e31158. Date of Electronic Publication: 2012 Feb 16. - Publication Year :
- 2012
-
Abstract
- Adhesion molecules play important roles in airway hyperresponsiveness or airway inflammation. Our previous study indicated catenin alpha-like 1 (CTNNAL1), an alpha-catenin-related protein, was downregulated in asthma patients and animal model. In this study, we observed that the expression of CTNNAL1 was increased in lung tissue of the ozone-stressed Balb/c mice model and in acute ozone stressed human bronchial epithelial cells (HBEC). In order to identify the possible DNA-binding proteins regulating the transcription of CTNNAL1 gene in HBEC, we designed 8 oligo- nucleotide probes corresponding to various regions of the CTNNAL1 promoter in electrophoretic mobility shift assays (EMSA). We detected 5 putative transcription factors binding sites within CTNNAL1 promoter region that can recruit LEF-1, AP-2α and CREB respectively by EMSA and antibody supershift assay. Chromatin immunoprecipitation (ChIP) assay verified that AP-2 α and LEF-1 could be recruited to the CTNNAL1 promoter. Therefore we further analyzed the functions of putative AP-2 and LEF-1 sites within CTNNAL1 promoter by site-directed mutagenesis of those sites within pGL3/FR/luc. We observed a reduction in human CTNNAL1 promoter activity of mutants of both AP-2α and LEF-1 sites. Pre-treatment with ASOs targeting LEF-1and AP-2α yielded significant reduction of ozone-stress-induced CTNNAL1 expression. The activation of AP-2α and LEF-1, followed by CTNNAL1 expression, showed a correlation during a 16-hour time course. Our data suggest that a robust transcriptional CTNNAL1 up-regulation occurs during acute ozone-induced stress and is mediated at least in part by ozone-induced recruitments of LEF-1 and AP-2α to the human CTNNAL1 promoter.
- Subjects :
- Animals
Binding Sites
DNA-Binding Proteins
Humans
Lymphoid Enhancer-Binding Factor 1 genetics
Mice
Ozone pharmacology
Promoter Regions, Genetic
Stress, Physiological genetics
Transcription Factor AP-2 genetics
Bronchi cytology
Epithelial Cells metabolism
Gene Expression Regulation
Lymphoid Enhancer-Binding Factor 1 metabolism
Transcription Factor AP-2 metabolism
Transcription Factors physiology
alpha Catenin genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 7
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 22359570
- Full Text :
- https://doi.org/10.1371/journal.pone.0031158