The DNA binding activity of metal response element-binding transcription factor-1 is activated in vivo and in vitro by zinc, but not by other transition metals

被引:164
作者
Bittel, D
Dalton, T
Samson, SLA
Gedamu, L
Andrews, GK
机构
[1] Univ Kansas, Med Ctr, Dept Biochem & Mol Biol, Kansas City, KS 66160 USA
[2] Univ Calgary, Dept Biol Sci, Calgary, AB T2N 1N4, Canada
关键词
D O I
10.1074/jbc.273.12.7127
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 examined the DNA binding activity of mouse and human MTF-1 in whole cell extracts from cells cultured in medium containing zinc or cadmium and from untreated cells after the in vitro addition of zinc or cadmium, as well as using recombinant MTF-1 transcribed and translated in vitro and treated with various transition metals, Incubation of human (HeLa) or mouse (Hepa) cells in medium containing cadmium (5-15 mu M) did not lead to a significant increase (<2-fold) in the amount of MTF-I DNA binding activity, whereas zinc (100 mu M) led to a 6-15-fold increase within 1 h, MTF-1 binding activity was low, but detectable, in control whole cell extracts and was increased (>10 fold) after the in vitro addition of zinc (30 mu M) and incubation at 37 degrees C for 15 min, In contrast, addition of cadmium (6 or 60 mu M) did not activate MTF-1 binding activity, Recombinant mouse and human MTF-1 were also dependent on exogenous zinc for DNA binding activity, Cadmium did not facilitate activation of recombinant MTF-1, but instead inhibited the activation of the recombinant protein by zinc, Interestingly, glutathione (1 mM) protected recombinant MIF-1 from inactivation by cadmium, and allowed for activation by zinc, It was also noted that zinc-activated recombinant MTF-1 was protected from cadmium only when bound to DNA These results suggest that cadmium interacts with the zinc fingers of MTF-1 and forms an inactive complex, Of the several transition metals (zinc, cadmium, nickel, silver, copper, and cobalt) examined, only zinc facilitated activation of the DNA binding activity of recombinant MTF-1, These data suggest that transition metals, other than zinc, that activate MT gene expression may do so by mechanisms independent of an increase in the DNA binding activity of MTF-1.
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收藏
页码:7127 / 7133
页数:7
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