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Journal of Bacteriology, March 2002, p. 1262-1269, Vol. 184, No. 5
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.5.1262-1269.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Reconstitution of the Trimethylamine Oxide Reductase Regulatory Elements of Shewanella oneidensis in Escherichia coli

*** Stéphanie Gon,1 Jean-Claude Patte,1 Jean-Philippe Dos Santos,1,{dagger} and Vincent Méjean1*

Laboratoire de Chimie Bactérienne, Institut de Biologie Structurale et Microbiologie, Centre National de la Recherche Scientifique, 13402 Marseille Cedex 20, France1

Received 20 September 2001/ Accepted 15 November 2001

Several bacteria can grow by using small organic compounds such as trimethylamine oxide (TMAO) as electron acceptors. In Shewanella species, the TMAO reductase respiratory system is encoded by the torECAD operon. We showed that production of the TMAO reductase of S. oneidensis was induced by TMAO and repressed by oxygen, and we noticed that a three-gene cluster (torSTR) encoding a complex two-component regulatory system was present downstream of the torECAD operon. We introduced the torSTR gene cluster into Escherichia coli and showed that this regulatory gene cluster is involved in TMAO induction of the torE promoter but plays no role in the oxygen control. The TorR response regulator was purified, and gel shift and footprinting experiments revealed that TorR binds to a single region located about 70 bases upstream of the transcription start site of the tor structural operon. By deletion analysis, we confirmed that the TorR operator site is required for induction of the tor structural promoter. As the TMAO regulatory system of S. oneidensis is homologous to that of E. coli, we investigated a possible complementation between the TMAO regulatory components of the two bacteria. Interestingly, TorSec, the TMAO sensor of E. coli, was able to transphosphorylate TorRso, the TMAO response regulator of S. oneidensis.

* Corresponding author. Mailing address: Laboratoire de Chimie Bactérienne, Institut de Biologie Structurale et Microbiologie, Centre National de la Recherche Scientifique, 31, chemin Joseph Aiguier, 13402 Marseille Cedex 20, France. Phone: (33) 4 91 16 40 32. Fax: (33) 4 91 71 89 14. E-mail: mejean{at}ibsm.cnrs-mrs.fr.

{dagger} Present address: HTS-BIO, Z.I. de Jouques, 13420 Gemenos, France.

Journal of Bacteriology, March 2002, p. 1262-1269, Vol. 184, No. 5
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.5.1262-1269.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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