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Journal of Bacteriology, December 2002, p. 6624-6634, Vol. 184, No. 23
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.23.6624-6634.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

The Nitrate Reductase and Nitrite Reductase Operons and the narT Gene of Staphylococcus carnosus Are Positively Controlled by the Novel Two-Component System NreBC

I. Fedtke, A. Kamps, B. Krismer, and F. Götz*

Mikrobielle Genetik, Universität Tübingen, 72076 Tübingen, Germany

Received 5 June 2002/ Accepted 24 August 2002

In Staphylococcus carnosus, the nreABC (for nitrogen regulation) genes were identified and shown to link the nitrate reductase operon (narGHJI) and the putative nitrate transporter gene narT. An nreABC deletion mutant, m1, was dramatically affected in nitrate and nitrite reduction and growth. Transcription of narT, narGHJI, and the nitrite reductase (nir) operon was severely reduced even when cells were cultivated anaerobically without nitrate or nitrite. nreABC transcripts were detected when cells were grown aerobically or anaerobically with or without nitrate or nitrite. NreA is a GAF domain-containing protein of unknown function. In vivo and in vitro studies showed that NreC is phosphorylated by NreB and that phospho-NreC specifically binds to a GC-rich palindromic sequence to enhance transcription initiation. This binding motif was found at the narGHJI, nir, and narT promoters but not at the moeB promoter. NreB is a cytosolic protein with four N-terminal cysteine residues. The second cysteine residue was shown to be important for NreB function. In vitro autophosphorylation of NreB was not affected by nitrate, nitrite, or molybdate. The nir promoter activity was iron dependent. The data provide evidence for a global regulatory system important for aerobic and anaerobic metabolism, with NreB and NreC forming a classical two-component system and NreB acting as a sensor protein with oxygen as the effector molecule.

* Corresponding author. Mailing address: Mikrobielle Genetik, Universität Tübingen, Waldhäuser Strasse 70-8, 72076 Tübingen, Germany. Phone: 49-7071-2974636. Fax: 49-7071-295937. E-mail: friedrich.goetz{at}uni-tuebingen.de.

Journal of Bacteriology, December 2002, p. 6624-6634, Vol. 184, No. 23
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.23.6624-6634.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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