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The function and structure of the metal coordination sites within the glucocorticoid receptor DNA binding domain

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Abstract

The glucocorticoid receptor enhances or represses transcription by binding to specific DNA sequences termed glucocorticoid response elements, or GREs1–5. Studies of cloned glucocorticoid receptors6–8 reveal that the protein is organized as functional domains, in an arrangement that appears to be common among members of the steroid receptor family9. A segment near the centre of the gene specifies DNA binding activity in vitro and contains two sequence motifs similar to 'zinc fingers' found in Xenopus transcription factor III A (TFIIIA)10. Such sequence motifs have been identified in nucleic acid binding proteins from a wide range of organisms11–13. Steroid receptor protein fingers are proposed to bind zinc through two pairs of conserved cysteine residues7,14,15.We report here that a protein of relative molecular mass 19,000 (Mr = 19K) encompassing the DNA-binding domain of the glucocorticoid receptor that has been overexpressed in Escherichia coli and purified to homogeneity reversibly ligates two Zn(II) or Cd(II) ions. We show that metal ions are required for specific DNA binding and proper folding. Using EXAFS (extended X-ray absorption fine structure) and visible light spectroscopies, we find that each Zn atom is coordinated in a tetrahedral arrangement by four cysteines.

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Author notes

  1. Ben F. Luisi, Ravi Basavappa and Paul B. Sigler: Department of Biochemistry & Molecular Biology, The University of Chicago, Chicago, Illinois 60637, USA

  2. Z. Richard Korszun: Department of Chemistry, University of Wisconsin-Parkside, Kenosha, Wisconsin 53141, USA

Authors and Affiliations

  1. Department of Biochemistry & Biophysics, University of California, San Francisco, California, 94143-0448, USA

    Leonard P. Freedman, Ben F. Luisi, Z. Richard Korszun, Ravi Basavappa, Paul B. Sigler & Keith R. Yamamoto

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  1. Leonard P. Freedman
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  2. Ben F. Luisi
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  3. Z. Richard Korszun
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  4. Ravi Basavappa
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  5. Paul B. Sigler
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  6. Keith R. Yamamoto
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Freedman, L., Luisi, B., Korszun, Z. et al. The function and structure of the metal coordination sites within the glucocorticoid receptor DNA binding domain. Nature 334, 543–546 (1988). https://doi.org/10.1038/334543a0

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