Metalloregulation of the tumor suppressor protein p53: zinc mediates the renaturation of p53 after exposure to metal chelators in vitro and in intact cells.
The tumor suppressor p53 is a transcription factor which binds DNA through a structurally complex domain stabilized by a zinc atom. Zinc chelation disrupts the architecture of this domain, inducing the protein to adopt an immunological phenotype identical to that of many mutant forms of p53. In this report, we used 65Zn to show that incorporation of zinc within the protein was required for folding in the 'wild-type' conformation capable of specific DNA-binding. Using a cellular assay, we show that addition of extracellular zinc at concentrations within the physiological range (5 microM) was required for renaturation and reactivation of wild-type p53. Among other divalent metals tested (Cd2+, Cu2+, Co2+, Fe2+ and Ni2+), only Co2+ at 125 microM had a similar effect. Recombinant metallothionein (MT), a metal chelator protein, was found to modulate p53 conformation in vitro. In cultured cells, overexpression of MT by transfection could modulate p53 transcriptional activity. Taken together, these results suggest that zinc binding plays a regulatory role in the control of p53 folding and DNA-binding activity.
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Group of Molecular Carcinogenesis, International Agency for Research on Cancer, 150 cours Albert Thomas, F-69372, Lyon Cedex 08, France.
Richard
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MJ
Hainaut
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D000818:Animals; D002104:Cadmium / Q000378:metabolism / Q000494:pharmacology; D002413:Cations, Divalent / Q000378:metabolism / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology*; D003035:Cobalt / Q000378:metabolism / Q000494:pharmacology; D004247:DNA / Q000235:genetics / Q000378:metabolism; D005029:Ethylenediamines / Q000494:pharmacology; D005347:Fibroblasts; D006801:Humans; D008668:Metallothionein / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects; D017510:Protein Folding; D020673:Protein Renaturation / Q000187:drug effects; D013816:Thermodynamics; D015533:Transcriptional Activation / Q000187:drug effects; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D015032:Zinc / Q000378:metabolism / Q000494:pharmacology*
D002413:Cations, Divalent; D002614:Chelating Agents; D005029:Ethylenediamines; D016159:Tumor Suppressor Protein p53; D002104:Cadmium; D003035:Cobalt; D004247:DNA; D008668:Metallothionein; D015032:Zinc; C044387:N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine