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Applied and Environmental Microbiology, August 2006, p. 5260-5265, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00599-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

The Bacillus subtilis nrdEF Genes, Encoding a Class Ib Ribonucleotide Reductase, Are Essential for Aerobic and Anaerobic Growth

Elisabeth Härtig,1* Anja Hartmann,1 Manuela Schätzle,2 Alessandra M. Albertini,3 and Dieter Jahn1

Institute of Microbiology, Technical University of Braunschweig, D-38106 Braunschweig, Germany,1 Division of Biophysical Chemistry, Biozentrum, University of Basel, Basel, Switzerland,2 Dipartimento di Genetica e Microbiologia, Università degli Studi di Pavia, Pavia, Italy3

Received 14 March 2006/ Accepted 31 May 2006

Ribonucleotide reductases (RNRs) are essential for the biosynthesis of the deoxyribonucleoside triphosphates of DNA. Recently, it was proposed that externally supplied deoxyribonucleosides or DNA is required for the growth of Bacillus subtilis under strict anaerobic conditions (M. J. Folmsbee, M. J. McInerney, and D. P. Nagle, Appl. Environ. Microbiol. 70:5252-5257, 2004). Cultivation of B. subtilis on minimal medium in the presence of oxygen indicators in combination with oxygen electrode measurements and viable cell counting demonstrated that growth occurred under strict anaerobic conditions in the absence of externally supplied deoxyribonucleosides. The nrdEF genes encode the only obvious RNR in B. subtilis. A temperature-sensitive nrdE mutant failed to grow under aerobic and anaerobic conditions, indicating that this oxygen-dependent class I RNR has an essential role under both growth conditions. Aerobic growth and anaerobic growth of the nrdE mutant were rescued by addition of deoxynucleotides. The nrd locus consists of an nrdI-nrdE-nrdF-ymaB operon. The 5' end of the corresponding mRNA revealed transcriptional start sites 45 and 48 bp upstream of the translational start of nrdI. Anaerobic transcription of the operon was found to be dependent on the presence of intact genes for the ResDE two-component redox regulatory system. Two potential ResD binding sites were identified approximately 62 bp (site A) and 50 bp (site B) upstream of the transcriptional start sites by a bioinformatic approach. Only mutation of site B eliminated nrd expression. Aerobic transcription was ResDE independent but required additional promoter elements localized between 88 and 275 bp upstream of the transcriptional start.

* Corresponding author. Mailing address: Institute of Microbiology, Technical University of Braunschweig, Spielmannstr. 7, D-38106 Braunschweig, Germany. Phone: 49 (0)531 391-5819. Fax: 49 (0)531 391-5854. E-mail: e.haertig{at}tu-bs.de.

Applied and Environmental Microbiology, August 2006, p. 5260-5265, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00599-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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