PUBLICATION
Cloning of zebrafish metallothionein gene and characterization of its gene promoter region in HepG2 cell line
- Authors
- Yan, C.H., and Chan, K.M.
- ID
- ZDB-PUB-040713-5
- Date
- 2004
- Source
- BBA Gene Structure and Expression 1679(1): 47-58 (Journal)
- Registered Authors
- Chan, King-Ming
- Keywords
- Heavy metal, Gene regulation, Electrophoretic mobility assay, Transcription factor
- MeSH Terms
-
- Amino Acid Sequence
- Animals
- Base Sequence
- Cell Line
- Cloning, Molecular
- DNA Primers
- Electrophoretic Mobility Shift Assay
- Humans
- Metallothionein/genetics*
- Molecular Sequence Data
- Promoter Regions, Genetic*
- Sequence Homology, Amino Acid
- Sequence Homology, Nucleic Acid
- Zebrafish
- PubMed
- 15245916 Full text @ BBA Gene Structure and Expression
- CTD
- 15245916
Citation
Yan, C.H., and Chan, K.M. (2004) Cloning of zebrafish metallothionein gene and characterization of its gene promoter region in HepG2 cell line. BBA Gene Structure and Expression. 1679(1):47-58.
Abstract
The coding region of cDNA and genomic DNA, with its promoter region, of zebrafish metallothionein (zMT) gene homologous to the piscine MT-II was obtained. The A/T-rich promoter region contains four metal regulatory elements (MREs), three activator protein 1 (AP1) and one specific protein 1 (Sp1) binding sites. The four MREs are organized into two clusters, a distal cluster with one MRE lying around 740 bp upstream of the transcription start point and a proximal cluster with three MREs located close to the TATA box. The metal induction ability of the promoter was assessed by transient luciferase gene expression assays in HepG2 cells. The zMT promoter was inducible by Zn(2+), Cd(2+), Cu(2+) and Hg(2+) ions in decreasing inducibility, while inert to Ni(2+), Pb(2+) and Co(2+) ions, and H(2)O(2) treatment in vitro. Deletion of the putative cis-acting elements in the promoter region revealed that the distal MRE (MREd) was important in mediating metal inducibility. Despite the binding of HepG2 cell nuclear protein factors to all MREs as confirmed by electrophoretic mobility shift assay (EMSA), the proximal MREs did not provide significant contribution to metal induction of zMT gene in HepG2 cells. The metal inducibility of zMT promoter required the cooperative effect of at least three MRE sites.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping