title: Characterization of non-inducible Tet repressor mutants suggests conformational changes necessary for induction.
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.
authors.affiliation: Lehrstuhl für Mikrobiologie, Friedrich-Alexander Universität Erlangen-Nürnberg, Germany.
authors.forename: G
authors.initials: G
authors.lastname: Müller
authors.forename: B
authors.initials: B
authors.lastname: Hecht
authors.forename: V
authors.initials: V
authors.lastname: Helbl
authors.forename: W
authors.initials: W
authors.lastname: Hinrichs
authors.forename: W
authors.initials: W
authors.lastname: Saenger
authors.forename: W
authors.initials: W
authors.lastname: Hillen
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013752:Tetracycline / Q000378:metabolism*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D046911:Macromolecular Substances; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C034811:tetracycline resistance-encoding transposon repressor protein; D004247:DNA; D001616:beta-Galactosidase; D013752:Tetracycline