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Base sequence discrimination by zinc-finger DNA-binding domains

Nature volume 349, pages 175–178 (1991)Cite this article

Abstract

ZINC fingers1,2 constitute important eukaryotic DNA-binding domains, being present in many transcription factors3–5. The Cys2/His2zinc-finger class has conserved motifs of 28–30 amino acids which are usually present as tandem repeats1,2. The structure of a Cys2/His2zinc finger has been determined by nuclear magnetic resonance6, but details of its interaction with DNA were not established. Here we identify amino acids governing DNA-binding specificity using in vitro directed mutagenesis guided by similarities between the zinc fingers of transcription factors Spl (ref. 7) and Krox-20 (ref. 8). Krox-20 is a serum-inducible transcription activator8,9 which is possibly involved in the regulation of hindbrain development10; it contains three zinc fingers similar to those of Spl (refs 7, 8, 11) and binds to a 9-base-pair target sequence which is related to that of Spl (ref. 9). Our results show that each finger spans three nucleotides and indicate two positions in Krox-20 zinc fingers that are important for base-pair selectivity. Modelling with molecular graphics suggests that these residues could bind directly with the bases and that other amino acid–base contacts are also possible.

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Authors and Affiliations

  1. Laboratoire de Génétique Moléculaire, CNRS D 1302, École Normale Supérieure, 46, rue d'Ulm, F-75230, Paris, Cedex 05, France

    Jeannette Nardelli, Christine Vesque & Patrick Charnay

  2. European Molecular Biology Laboratory, Meyerhofstrasse 1, Postfach 10-2209, D-6900, Heidelberg, Germany

    Toby J. Gibson

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  1. Jeannette Nardelli
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  2. Toby J. Gibson
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  3. Christine Vesque
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  4. Patrick Charnay
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Nardelli, J., Gibson, T., Vesque, C. et al. Base sequence discrimination by zinc-finger DNA-binding domains. Nature 349, 175–178 (1991). https://doi.org/10.1038/349175a0

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