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Molecular cloning and characterization of human acid sensing ion channel (ASIC)2 gene promoter
- Source :
- Gene. 313
- Publication Year :
- 2003
-
Abstract
- Acid sensing ion channel (ASIC)2 belongs to the amiloride-sensitive Na(+)-channel/ degenerin family. Our previous studies suggested that differential regulation of ASIC2 expression occurs between high-grade glial-derived tumor cells and normal astrocytes. To investigate the mechanisms involved in the regulation of ASIC2 gene expression, the human ASIC2 promoter region (-1551 to +117) was cloned and characterized. The ASIC2 promoter lacked a canonical TATA box, but contained one putative CCAAT box. Nucleotide sequencing of the promoter revealed the presence of a number of transcription factor-binding sites and a 404 bp CpG island upstream the transcription start site. Nested deletion mutants and transfection results showed that the construct between -133 and +117 base pairs conferred basal transcription specific activity. Mutation of Sp1 and CP2 sites in this region resulted in a 70 and 95% decrease in promoter activity, respectively. Gel shift assays demonstrated the existence of specific protein binding to the SP1 and CP2 elements. There was no mutation in the CpG island in six glioma cell lines, but methylation-specific PCR showed methylation in some of glioma cell lines and tumor tissues, and treatment with the methylation inhibitor 5-Aza-2'-deoxycytidine could partially restore ASIC2 expression in cell lines, suggesting that epigenetic mechanisms may contribute to dysregulated ASIC2 expression.
- Subjects :
- Patch-Clamp Techniques
Sp1 Transcription Factor
TATA box
Recombinant Fusion Proteins
Molecular Sequence Data
CAAT box
Nerve Tissue Proteins
Biology
Decitabine
Transfection
Sodium Channels
Membrane Potentials
Epigenetics of physical exercise
Genetics
Tumor Cells, Cultured
Humans
RNA, Messenger
Cloning, Molecular
Luciferases
Promoter Regions, Genetic
Transcription factor
Regulation of gene expression
Binding Sites
General transcription factor
Base Sequence
Brain
Membrane Proteins
RNA-Binding Proteins
Promoter
General Medicine
DNA
Glioma
Sequence Analysis, DNA
DNA Methylation
Molecular biology
Acid Sensing Ion Channels
DNA-Binding Proteins
Gene Expression Regulation, Neoplastic
DNA methylation
Mutation
Azacitidine
CpG Islands
Transcription Initiation Site
Transcription Factors
Subjects
Details
- ISSN :
- 03781119
- Volume :
- 313
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....dc9686c98f6637cd2d164436305a40f2