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Characterization of non-inducible Tet repressor mutants suggests conformational changes necessary for induction

Nature Structural Biology volume 2, pages 693–703 (1995)Cite this article

Abstract

Non-inducible tetracycline repressor (TetR) mutants were grouped in three structurally distinct classes. We quantitated in vivo operator binding, inducibility, and in vitro tetracycline binding of mutants from each class. Mutation of residues close to tetracycline (class 1) leads to reduced affinity for the drug. Mutation of residues located at the connection of the DNA-reading head with the protein core (class 2) and at the dimerization interface (class 3) bind inducer with the same affinity as wild-type TetR. These mutations interfere with the induced, but not the operator-binding conformation of TetR. The affinity of some class 1 mutants for tetracycline is less affected than their inducibility, suggesting that the mutated residues are important for triggering those conformational changes necessary for induction.

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Authors and Affiliations

  1. Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie, Biochemie und Genetik der Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstr. 5, 91058, Erlangen, Germany

    Gerhard Müller, Brigitte Hecht, Vera Helbl & Wolfgang Hillen

  2. Institut für Kristallographie, Freie Universität Berlin, Takusstr. 6, 14195, Berlin, Germany

    Winfried Hinrichs & Wolfram Saenger

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  1. Gerhard Müller
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Müller, G., Hecht, B., Helbl, V. et al. Characterization of non-inducible Tet repressor mutants suggests conformational changes necessary for induction. Nat Struct Mol Biol 2, 693–703 (1995). https://doi.org/10.1038/nsb0895-693

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