title: TorT, a member of a new periplasmic binding protein family, triggers induction of the Tor respiratory system upon trimethylamine N-oxide electron-acceptor binding in Escherichia coli.
abstract: In anaerobiosis, Escherichia coli can use trimethylamine N-oxide (TMAO) as a terminal electron acceptor. Reduction of TMAO in trimethylamine (TMA) is mainly performed by the respiratory TMAO reductase. This system is encoded by the torCAD operon, which is induced in the presence of TMAO. This regulation involves a two-component system comprising TorS, an unorthodox histidine kinase, and TorR, a response regulator. A third protein, TorT, sharing homologies with periplasmic binding proteins, plays a key role in this regulation because disruption of the torT gene abolishes tor expression. In this study we showed that TMAO protects TorT against degradation by the GluC endoproteinase and modifies its temperature-induced CD spectrum. We also isolated a TorT negative mutant that is no longer protected by TMAO from degradation by GluC. Isothermal titration calorimetry confirmed that TorT binds TMAO with a binding constant of 150 mum. Therefore, we conclude that TorT binds TMAO and that this binding promotes a conformational change of TorT. We also showed that TorT interacts with the periplasmic domain of TorS in both the presence and absence of TMAO but the TorT-TMAO complex induces a higher GluC protection of TorS than TorT alone. These results support the idea that TMAO binding to TorT induces a cascade of conformational changes from TorT to TorS, leading to TorS activation. We identified several homologues of the TorT protein that define a new family of periplasmic binding proteins. We thus propose that the members of this family bind TMAO or related compounds and that they are involved in signal transduction or even substrate transport.
authors.affiliation: Laboratoire de Chimie Bactérienne, Laboratoire de Bioénergétique et Ingénierie des Protéines, Institut de Biologie Structurale et Microbiologie, Centre National de la Recherche Scientifique, 31 chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
authors.forename: Claudine
authors.initials: C
authors.lastname: Baraquet
authors.forename: Laurence
authors.initials: L
authors.lastname: Théraulaz
authors.forename: Marianne
authors.initials: M
authors.lastname: Guiral
authors.forename: Daniel
authors.initials: D
authors.lastname: Lafitte
authors.forename: Vincent
authors.initials: V
authors.lastname: Méjean
authors.forename: Cécile
authors.initials: C
authors.lastname: Jourlin-Castelli
mesh_terms: D000595:Amino Acid Sequence; D002151:Calorimetry; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D013058:Mass Spectrometry; D008744:Methylamines / Q000378:metabolism*; D008969:Molecular Sequence Data; D033903:Periplasmic Proteins / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D010957:Plasmids; D011485:Protein Binding; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
chemical_list: D029968:Escherichia coli Proteins; D008744:Methylamines; D033903:Periplasmic Proteins; C098062:TorT protein, E coli; C005855:trimethyloxamine