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Journal of Bacteriology, March 1999, p. 1481-1488, Vol. 181, No. 5
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Molecular Characterization of the Nitrite-Reducing System of Staphylococcus carnosus

H. Neubauer,dagger I. Pantel, and F. Götz*

Mikrobielle Genetik, Universität Tübingen, 72076 Tübingen, Germany

Received 15 January 1998/Accepted 10 December 1998

Characterization of a nitrite reductase-negative Staphylococcus carnosus Tn917 mutant led to the identification of the nir operon, which encodes NirBD, the dissimilatory NADH-dependent nitrite reductase; SirA, the putative oxidase and chelatase, and SirB, the uroporphyrinogen III methylase, both of which are necessary for biosynthesis of the siroheme prosthetic group; and NirR, which revealed no convincing similarity to proteins with known functions. We suggest that NirR is essential for nir promoter activity. In the absence of NirR, a weak promoter upstream of sirA seems to drive transcription of sirA, nirB, nirD, and sirB in the stationary-growth phase. In primer extension experiments one predominant and several weaker transcription start sites were identified in the nir promoter region. Northern blot analyses indicated that anaerobiosis and nitrite are induction factors of the nir operon: cells grown aerobically with nitrite revealed small amounts of full-length transcript whereas cells grown anaerobically with or without nitrite showed large amounts of full-length transcript. Although a transcript is detectable, no nitrite reduction occurs in cells grown aerobically with nitrite, indicating an additional oxygen-controlled step at the level of translation, enzyme folding, assembly, or insertion of prosthetic groups. The nitrite-reducing activity expressed during anaerobiosis is switched off reversibly when the oxygen tension increases, most likely due to competition for electrons with the aerobic respiratory chain. Another gene, nirC, is located upstream of the nir operon. nirC encodes a putative integral membrane-spanning protein of unknown function. A nirC mutant showed no distinct phenotype.

* 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.

dagger Present address: Centre de Recherche Nestlé, Biologie Moleculaire, Vers-chez-les Blanc, CH-1000 Lausanne 26, Switzerland.

Journal of Bacteriology, March 1999, p. 1481-1488, Vol. 181, No. 5
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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