THE TRANSCRIPTION FACTOR MTF-1 IS ESSENTIAL FOR BASAL AND HEAVY METAL-INDUCED METALLOTHIONEIN GENE-EXPRESSION

被引:412
作者
HEUCHEL, R
RADTKE, F
GEORGIEV, O
STARK, G
AGUET, M
SCHAFFNER, W
机构
[1] UNIV ZURICH, INST MOLEK BIOL 2, CH-8057 ZURICH, SWITZERLAND
[2] UNIV ZURICH, INST MOLEK BIOL 1, CH-8093 ZURICH, SWITZERLAND
关键词
METALLOTHIONEIN GENE EXPRESSION; TARGETED GENE DISRUPTION; TRANSCRIPTION FACTOR MTF-1; ZINC FINGER PROTEIN;
D O I
10.1002/j.1460-2075.1994.tb06581.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have described and cloned previously a factor (MTF-1) that binds specifically to heavy metal-responsive DNA sequence elements in the enhancer/promoter region of metallothionein genes. MTF-1 is a protein of 72.5 kDa that contains six zinc fingers and multiple domains for transcriptional activation. Here we report the disruption of both alleles of the MTF-1 gene in mouse embryonic stem cells by homologous recombination. The resulting null mutant cell line fails to produce detectable amounts of MTF-1. Moreover, due to the loss of MTF-1, the endogenous metallothionein I and II genes are silent, indicating that MTF-1 is required for both their basal and zinc-induced transcription. In addition to zinc, other heavy metals, including cadmium, copper, nickel and lead, also fail to activate metal-responsive promoters in null mutant cells. However, cotransfection of an MTF-1 expression vector and metal-responsive reporter genes yields strong basal transcription that can be further boosted by zinc treatment of cells. These results demonstrate that MTF-1 is essential for metallothionein gene regulation. Finally, we present evidence that MTF-1 itself is a zinc sensor, which exhibits increased DNA binding activity upon zinc treatment.
引用
收藏
页码:2870 / 2875
页数:6
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