Isolation and characterization of the human CLC-5 chloride channel gene promoter
| Autor: | Atsushi Hayama, Fumiaki Marumo, Shinichi Uchida, Sei Sasaki |
|---|---|
| Rok vydání: | 2000 |
| Předmět: |
DNA
Complementary Transcription Genetic RNA Splicing Recombinant Fusion Proteins TATA box Molecular Sequence Data CHO Cells Biology medicine.disease_cause Cell Line Exon Chloride Channels Cricetinae Sequence Homology Nucleic Acid Complementary DNA Cyclic AMP Genetics medicine Animals Humans Cloning Molecular Luciferases Promoter Regions Genetic Gene Sequence Deletion Mutation Binding Sites Dent's disease Base Sequence Dose-Response Relationship Drug urogenital system CLCN5 Colforsin Promoter DNA Sequence Analysis DNA General Medicine Thionucleotides medicine.disease Molecular biology Transcription Factor AP-1 Gene Expression Regulation Parathyroid Hormone biology.protein Tetradecanoylphorbol Acetate Protein Binding |
| Zdroj: | Gene. 261:355-364 |
| ISSN: | 0378-1119 |
| DOI: | 10.1016/s0378-1119(00)00493-5 |
| Popis: | The human CLC-5 chloride channel is expressed mainly in the kidney and its mutations cause Dent's disease (a familial renal tubular syndrome with hypercalciuria, tubular proteinuria, rickets, nephrocalcinosis, and eventual renal failure). To gain insight into the regulatory mechanism of CLC-5 expression, a genomic clone that contains the 5'-flanking region of the human CLC-5 gene was isolated and characterized. Two types of 5'-ends of cDNA were isolated by 5'-rapid amplification of cDNA ends, and one of them, approximately 2.1 kbp upstream of ATG-containing exon II, was first identified in human. The major promoter activity was detected in the 5'-flanking region of this newly identified exon Ia. The sequence of the proximal 5'-flanking region contained an activator protein (AP)-1-like site and cAMP-responsive element, but it lacked a TATA box, a GC-rich element, and an SP-1 site. Deletion analysis of the 5'-flanking region showed that the fragments containing the AP-1-like element (TGACTCC) positioned at -38 exhibited high promoter activities in CLC-5 expressing LLC-PK1 cells, but that further deletions not containing this AP-1-like element resulted in a great loss of luciferase activities. Gel-retardation analysis demonstrated the existence of a specific protein binding to this AP-1-like element in LLC-PK1 cells, which seemed to differ from an authentic AP-1. This study clarified the key element of the human CLCN5 promoter, and the mutation in this region could be the cause of Dent's disease. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect