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Nucleic Acids Research, 1988, Vol. 16, No. 22 10547-10560
© 1988

MOLECULAR BIOLOGY

Detection of a nuclear protein that interacts with a metal regulatory element of the mouse metallothionein 1 gene

Carl Séguin*,1 and Jacinthe Prévost1

Department of Physiology and Laval University Medical Center, Laval University Québec, Canada

*To whom correspondence should be addressed

Received August 15, 1988. Revised October 21, 1988. Accepted October 21, 1988.

Metallothionein (MT) genes contain multiple metal regulatory elements (MREs) that are responsible for metal induction. A protein blotting procedure and a synthetic oligonucleotide have been used to identify nuclear factors interacting with a MRE (MREd) of the mouse MT-1 gene. We report the specific binding of the probe to a protein of apparent Mr 108,000 (p108). The specificity of the interaction was demonstrated by mutation analysis and competition experiments. Furthermore, the probe contains the Sp1 consensus binding sequence 5'CCGCCC3', in addition to the MRE consensus sequence, 5'TGCAC3', and we show that a Simian Virus 40 DNA fragment which contains six Sp1 binding sites did not bind p108 nor did it compete for the protein(s) interacting with MREd in a DNA footprinting assay. These results show that a metal regulatory element of the mouse MT-1 gene interacts specifically with a nuclear protein of Mr 108,000 and that this protein is distinct from the transcription factor Sp1.

1Present address: Centre de Recherches en Cancérologie de l'Université Laval, L'Hôtel-Dieu de Québec, 11 Côte du Palais, Québec, Qc, G1R 2J6, Canada.


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