Effect of copper, zinc and cadmium on the promoter of selenoprotein W in glial and myoblast cells.
Rat selenoprotein W (SeW) promoter activity was investigated using different concentrations of cadmium, copper, and zinc. Two fragments (404 and 1265 bp) of the SeW promoter, containing a single metal response element (MRE), were ligated into the multiple cloning site of a pGL3-Basic reporter plasmid. The constructs were transfected into cultured C6 (rat glial) and L8 (myoblast) cells and promoter activity measured by means of luciferase reporter gene fused to the SeW promoter fragments in the reporter plasmid. With post-transfection exposure of these cell lines to these metals, copper and zinc, but not cadmium, significantly increased promoter activity of the unmutated 1265 bp (not 404 bp) construct (p<0.05) only in the C6 cells. Mutation of the MRE sequence abolished promoter response to metal exposure but did not eliminate promoter activity. The results suggest that SeW expression in glial cells can be increased on exposure to copper and zinc and that this response is dependent on the MRE sequence present in the SeW promoter.
Amantana
A
A
Department of Environmental and Molecular Toxicology, Agricultural and Life Sciences 1007, Oregon State University, Corvallis, OR 97331-7301, USA.
Vorachek
W R
WR
Butler
J A
JA
Costa
N D
ND
Whanger
P D
PD
D000818:Animals; D001483:Base Sequence; D002104:Cadmium / Q000494:pharmacology*; D003300:Copper / Q000494:pharmacology*; D004247:DNA; D008969:Molecular Sequence Data; D009132:Muscles / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D009457:Neuroglia / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011506:Proteins / Q000235:genetics*; D051381:Rats; D051151:Selenoprotein W; D051140:Selenoproteins; D015032:Zinc / Q000494:pharmacology*
D011506:Proteins; D051151:Selenoprotein W; D051140:Selenoproteins; D002104:Cadmium; D003300:Copper; D004247:DNA; D015032:Zinc