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Title: Retrovirally expressed metal response element-binding transcription factor-1 normalizes metallothionein-1 gene expression and protects cells against zinc, but not cadmium, toxicity.

Authors: Solis, Willy A; Childs, Nicole L; Weedon, Michael N; He, Lei; Nebert, Daniel W; Dalton, Timothy P

Published In Toxicol Appl Pharmacol, (2002 Jan 15)

Abstract: Metal response element (MRE) transcription factor-1 (MTF1), a member of the Cys2-His2 class of zinc-finger transcription factors, is best known for its robust transcriptional regulation of mammalian metallothionein (MT) genes. MTF1 is also believed to play a generalized role in regulating genes involved in protection against heavy metals and oxidative stress. MTF1 binding to MRE motifs is regulated by changes in intracellular zinc (Zn(2+)) concentration. Molecular dissection of MTF1 has been hindered by its high constitutive trans-activity following transient transfection and the failure of these systems to examine genes packaged in native chromatin. In developing a system to avoid these problems, we employed a high-efficiency retroviral transduction system to reintroduce MTF1 into mouse Mtf1(-/-) knockout cells (dko7). Electrophoretic mobility shift assays demonstrated that MTF1 retrovirally transduced dko7 cells (MTF1dko7) possess levels of inducible MTF1-MRE binding activity similar to that seen in mouse hepatoma Hepa-1 cells, and MTF1 binding could be modulated over a 20-fold range by varying the concentration of Zn(2+) present in the culture medium. The dko7 cells exhibited no change in Mt1 gene expression upon Zn(2+) or cadmium (Cd(2+)) treatment; in contrast, in MTF1dko7 cells, Zn(2+) or Cd(2+) induced MT1 mRNA accumulation in a dose-dependent manner. Interestingly, MTF1dko7 cells showed resistance to Zn(2+) toxicity, but negligible resistance to Cd(2+). Concomitantly, MT1 protein levels in MTF1dko7 cells were inducible to the same degree as that in Hepa-1 cells when treated with Zn(2+), but not with Cd(2+). Together, our studies suggest that MTF1-mediated regulation of gene expression is sufficient to protect cells against Zn(2+) toxicity and may be necessary but not sufficient to protect cells against Cd(2+) toxicity.

PubMed ID: 11814329 Exiting the NIEHS site

MeSH Terms: Animals; Cadmium/antagonists & inhibitors*; Cadmium/toxicity*; Cell Survival/drug effects; DNA, Complementary/biosynthesis; DNA, Complementary/genetics; Electrophoresis; Gene Expression Regulation*; Genetic Vectors; In Situ Hybridization; Liver Neoplasms, Experimental/genetics; Liver Neoplasms, Experimental/metabolism; Metallothionein/biosynthesis*; Metallothionein/genetics*; Mice; Mice, Knockout; Plasmids/genetics; RNA, Messenger/biosynthesis; RNA, Messenger/genetics; Research Support, U.S. Gov't, P.H.S.; Retroviridae*; Transcription Factors/biosynthesis; Transcription Factors/genetics*; Transduction, Genetic; Tumor Cells, Cultured; Zinc/antagonists & inhibitors*; Zinc/toxicity*

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