Abstract
We explored the molecular genetics underlying the massive induction of isoMTs by Zn2+ or Cd2+ in metal tolerant rabbit kidney (RK-13) sub-line cells, using band shift assays and Southern blotting analysis. In sub-line cells accommodated to intermediate metal concentrations (100 μM Zn2+; 1–20 μM Cd2+) evidence suggested that the increase in the capacity for isoMT synthesis is brought about by an increased binding activity of the nuclear transcription factors MTF-1 and Sp1. Using quantitative band shift analysis with a mouse MRE-d oligonucleotide probe, the binding of both transcription factors was found to be enhanced two to three times over the binding activity measured in the unexposed parental RK-13 cells. Their increase in binding activity is probably the cause of the overexpression of MT genes and the development of metal tolerance in these cells. In cells tolerant to the highest concentrations of metal the analysis of Southern blot signals revealed MT gene amplification to be the most probable cause of the increased MT production. Thus, in cells of sub-lines growing in the presence of 350 μM Zn2+, two of the isoMT genes were coordinately triplicated and in cells tolerant to 150 μM Cd2+ one isoMT gene was amplified two-fold.
Similar content being viewed by others
References
Carter, A. D., Felber, B. K., Walling, M. J., Jubier, M.-F., Schmidt, C. J., and Hamer, D. H., Proc. natl Acad. Sci. USA81 (1984) 7392.
Karin, M., Haslinger, H., Richards, R. I., Krauter, P., Westphal, H. M., and Beato, M., Nature, Lond.308 (1984) 513.
Stuart, G. W., Searle, P. F., Chen, H. Y., Brinster, R. I., and Palmiter, R. D., Proc. natl Acad. Sci. USA81 (1984) 7318.
Seguin, C., Felber, B. K., Carter, A. D., and Hamer, D. H., Nature Lond.312 (1984) 781.
Westin, G., and Schaffner, W., EMBO J.7 (1988) 3763.
Beach, L. R., and Palmiter, R. D., Proc. natl Acad. Sci. USA78 (1981) 2110.
Gick, G. G., and McCarty, K. S., J. biol. Chem.257 (1982) 9049.
Wan, M., Hunziker, P. E., and Kägi, J. H. R., Biochem. J.292 (1993) 609.
Hamer, D. H., A. Rev. Biochem.55 (1986) 913.
Palmiter, R. D., in: Metallothionein II, p. 63. Eds J. H. R. Kägi and Y. Kojima. Birkhäuser, Basel 1987.
Koropatnick, J., Proc. Soc. expl Biol. Med.188 (1988) 287.
Foster, R., and Gedamu, L., J. biol. Chem.266 (1991) 9866.
Schreiber, E., Matthias, P., Müller, M. M., and Schaffner, W., Nucleic Acids Res.7 (1989) 6419.
Kemler, I., Schreiber, E., Müller, M. M., Matthias, P., and Schaffner, W., EMBO J.8 (1989) 2001.
Sambrook, J., Fritsch, E. F., and Maniatis, T., Molecular Cloning. Cold Spring Harbor, New York 1989.
Tam, Y. C., Chopra, A., Hassan, M., and Thirion, J. P., Biochem. biophys. Res. Commun.153 (1988) 209.
Seguin, C., and Hamer, D. H., Science235 (1987) 1383.
Schöler, H., Haslinger, A., Heguy, A., and Karin, M., Science232 (1986) 76.
Andersen, R. D., Taplitz, S. J., Wong, S., Bristol, G., Larkin, B., and Herschman, H. R., Molec. cell. Biol.7 (1987) 3574.
Koropatnick, J., Winning, R., Wiese, E., Heschl, M., Gedamu, L., and Duerksen, J., Nucleic Acids Res.13 (1985) 5423.
Palmiter, R. D., Proc. natl Acad. Sci. USA91 (1994) 1219.
Author information
Authors and Affiliations
Additional information
A full list of abbreviations is given at the end of the article.
Rights and permissions
About this article
Cite this article
Wan, M., Heuchel, R., Radtke, F. et al. Regulation of metallothionein gene expression in Cd- or Zn-adapted RK-13 cells. Experientia 51, 606–611 (1995). https://doi.org/10.1007/BF02128753
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF02128753