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J. Biol. Chem., Vol. 275, Issue 44, 34803-34809, November 3, 2000
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From the Department of Biochemistry & Molecular Biology, University
of Kansas Medical Center, Kansas City, Kansas 66160
Metal regulation of the mouse zinc transporter
(ZnT)-1 gene was examined in cultured cells and in the developing
conceptus. Zinc or cadmium treatment of cell lines rapidly (3 h) and
dramatically (about 12-fold) induced ZnT1 mRNA levels. In cells
incubated in medium supplemented with Chelex-treated fetal bovine
serum, to remove metal ions, levels of ZnT1 mRNA were reduced, and
induction of this message in response to zinc or cadmium was
accentuated (up to 31-fold induction). Changes in ZnT1 gene expression
in these experiments paralleled those of metallothionein I (MT-I). Inhibition of RNA synthesis blocked metal induction of ZnT1 and MT-I
mRNAs, whereas inhibition of protein synthesis did not. Metal response element-binding transcription factor (MTF)-1 mediates metal
regulation of the metallothionein I gene. In vitro
DNA-binding assays demonstrated that mouse MTF-1 can bind avidly to the
two metal-response element sequences found in the ZnT1 promoter. Using mouse embryo fibroblasts with homozygous deletions of the MTF-1 gene,
it was shown that this transcription factor is essential for basal as
well as metal (zinc and cadmium) regulation of the ZnT1 gene in these
cells. In vivo, ZnT1 mRNA was abundant in the midgestation visceral yolk sac and placenta. Dietary zinc deficiency during pregnancy down-regulated ZnT1 and MT-I mRNA levels
(4-5-fold and >20-fold, respectively) in the visceral yolk sac, but
had little effect on these mRNAs in the placenta. Homozygous
knockout of the MTF-1 gene in transgenic mice also led to a 4-6-fold
reduction in ZnT1 mRNA levels and a loss of MT-I mRNA in the
visceral yolk sac. These results suggest that MTF-1 mediates the
response to metal ions of both the ZnT1 and the MT-I genes the visceral
yolk sac. Overall, these studies suggest that MTF-1 directly
coordinates the regulation of genes involved in zinc homeostasis and
protection against metal toxicity.
The Transcription Factor MTF-1 Mediates Metal Regulation of
the Mouse ZnT1 Gene*
*
This work was supported by National Institutes of Health
Grant CA 61262 (to G. K. A.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence and reprint requests should be addressed:
Dept. of Biochemistry & Molecular Biology, University of Kansas Medical
Center, 3901 Rainbow Blvd., Kansas City, KS 66160-7421: Tel.:
913-588-6935; Fax: 913-588-7035; E-mail:
gandrews@kumc.edu.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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