Cloning of crucian carp ( Carassius cuvieri) metallothionein-II gene and characterization of its gene promoter region

The genomic DNA of crucian carp ( Carassius cuvieri) metallothionein-II (ccMT-II), with its upstream region, was obtained. The sequence analysis of its upstream region revealed several putative cis-acting elements including seven metal regulatory elements (MREs), three activator protein 1 (AP1), two...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 2006-04, Vol.342 (4), p.1297-1304
Main Authors: Ren, Hongwei, Xu, Mingxu, He, Pengfei, Muto, Norio, Itoh, Norio, Tanaka, Keiichi, Xing, Juan, Chu, Mingming
Format: Article
Language:English
Subjects:
Animals
Base Sequence
Carassius cuvieri
Carps - genetics
Cells, Cultured
Cloning, Molecular
Gene regulation
GRE
Metallothionein
Metallothionein - genetics
Molecular Sequence Data
MRE
Promoter Regions, Genetic - genetics
Sequence Homology, Nucleic Acid
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Summary:The genomic DNA of crucian carp ( Carassius cuvieri) metallothionein-II (ccMT-II), with its upstream region, was obtained. The sequence analysis of its upstream region revealed several putative cis-acting elements including seven metal regulatory elements (MREs), three activator protein 1 (AP1), two glucocorticoid response elements (GREs), etc. The seven MREs locate into two clusters, a distal cluster with four MREs within −800/−600 bp from the translation start site and a proximal cluster with three MREs close to TATA box. In transient luciferase gene expression assays, both of the distal and proximal cluster MREs have significantly shown synergistic effects in the transcription of ccMT-II gene; the proximal cluster of MREs serves as the major elements in metal inducing activity; Zn 2+ and Cd 2+ served as much stronger inducers than Cu 2+ shown in ccMT-II expression. The two GRE homologous sequences in ccMT-II promoter showed not to be inductive in either HepG2 or HEK293.
ISSN:0006-291X
1090-2104
DOI:10.1016/j.bbrc.2006.02.082