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Journal of Bacteriology, December 2001, p. 6815-6821, Vol. 183, No. 23
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.23.6815-6821.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Modulation of Anaerobic Energy Metabolism of Bacillus subtilis by arfM (ywiD)

Marco Marino,^1,2 Hugo Cruz Ramos,^3, Tamara Hoffmann,^1,4 Philippe Glaser,³ and Dieter Jahn^1,2,*

Institut für Organische Chemie und Biochemie, Albert-Ludwigs-Universität Freiburg, 79104 Freiburg,¹ Institut für Mikrobiologie, Technische Universität Braunschweig, 38106 Braunschweig,² and Max-Planck-Institut für Terrestrische Mikrobiologie, 35043 Marburg, and Laboratorium für Mikrobiologie, Fachbereich Biologie, Philipps-Universität Marburg, 35032 Marburg,⁴ Germany, and Unité de Régulation de l'Expression Génétique, Institut Pasteur, 75724 Paris Cedex 15, France³

Received 23 April 2001/Accepted 23 August 2001

Bacillus subtilis grows under anaerobic conditions utilizing nitrate ammonification and various fermentative processes. The two-component regulatory system ResDE and the redox regulator Fnr are the currently known parts of the regulatory system for anaerobic adaptation. Mutation of the open reading frame ywiD located upstream of the respiratory nitrate reductase operon narGHJI resulted in elimination of the contribution of nitrite dissimilation to anaerobic nitrate respiratory growth. Significantly reduced nitrite reductase (NasDE) activity was detected, while respiratory nitrate reductase activity was unchanged. Anaerobic induction of nasDE expression was found to be significantly dependent on intact ywiD, while anaerobic narGHJI expression was ywiD independent. Anaerobic transcription of hmp, encoding a flavohemoglobin-like protein, and of the fermentative operons lctEP and alsSD, responsible for lactate and acetoin formation, was partially dependent on ywiD. Expression of pta, encoding phosphotransacetylase involved in fermentative acetate formation, was not influenced by ywiD. Transcription of the ywiD gene was anaerobically induced by the redox regulator Fnr via the conserved Fnr-box (TGTGA-6N-TCACT) centered 40.5 bp upstream of the transcriptional start site. Anaerobic induction of ywiD by resDE was found to be indirect via resDE-dependent activation of fnr. The ywiD gene is subject to autorepression and nitrite repression. These results suggest a ResDE  Fnr  YwiD regulatory cascade for the modulation of genes involved in the anaerobic metabolism of B. subtilis. Therefore, ywiD was renamed arfM for anaerobic respiration and fermentation modulator.

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^* Corresponding author. Mailing address: Institut für Mikrobiologie, Technische Universität Braunschweig, Spielmannstr.7, 38106 Braunschweig, Germany. Phone: 49(0)531-3915801. Fax: 49(0)531-3915854. E-mail: d.jahn{at}tu-bs.de.

Present address: Department of Molecular Biology and Biotechnology, The University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, United Kingdom.

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Journal of Bacteriology, December 2001, p. 6815-6821, Vol. 183, No. 23
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.23.6815-6821.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

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