title: A peptide triggers allostery in tet repressor by binding to a unique site.
abstract: Regulatory proteins often communicate with each other to manage various cellular processes. Such interactions mostly rely on the recognition of small peptide motifs. The activity of other regulatory proteins depends on small molecular weight effectors and allostery. We demonstrate the in vivo regulation of the tetracycline-dependent Tet repressor by an oligopeptide fused to the N or C terminus of thioredoxin A. The binding site of the peptide overlaps but is not identical with the tetracycline binding site. Several TetR mutants that are non-inducible by tetracycline also respond to the peptide. This demonstrates for the first time the conversion of a small molecular weight effector-dependent regulator to a protein-protein contact-dependent potential member of designed signaling chains.
authors.affiliation: Lehrstuhl für Mikrobiologie, Institut für Mikrobiologie, Biochemie und Genetik, Friedrich-Alexander Universität Erlangen-Nürnberg, Staudtstrasse 5, 91058 Erlangen, Germany.
authors.forename: Marcus
authors.initials: M
authors.lastname: Klotzsche
authors.forename: Christian
authors.initials: C
authors.lastname: Berens
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hillen
mesh_terms: D000495:Allosteric Site; D000595:Amino Acid Sequence; D001665:Binding Sites; D015153:Blotting, Western; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D007700:Kinetics; D008954:Models, Biological; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D009842:Oligopeptides / Q000737:chemistry; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013753:Tetracycline Resistance; D013879:Thioredoxins / Q000737:chemistry; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D029968:Escherichia coli Proteins; D009842:Oligopeptides; D019151:Peptide Library; D010455:Peptides; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C470187:TetR(B) protein, E coli; C521715:pepBs1-Ac peptide; C034811:tetracycline resistance-encoding transposon repressor protein; D007544:Isopropyl Thiogalactoside; D013879:Thioredoxins; D001616:beta-Galactosidase