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Journal of Bacteriology, October 2004, p. 6477-6484, Vol. 186, No. 19
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.19.6477-6484.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Bacillus subtilis ResD Induces Expression of the Potential Regulatory Genes yclJK upon Oxygen Limitation

Elisabeth Härtig,1,{dagger}* Hao Geng,{dagger},2 Anja Hartmann,1 Angela Hubacek,3 Richard Münch,1 Rick W. Ye,4 Dieter Jahn,1 and Michiko M. Nakano2

Institute of Microbiology, University of Braunschweig, Braunschweig, Germany,1 Department of Environmental and Biomolecular Systems, OGI School of Science and Engineering, Oregon Health and Science University, Beaverton,2 Woodburn High School, Woodburn Oregon,3 Experimental Station E328/148B, DuPont Central Research and Development, Wilmington, Delaware4

Received 17 May 2004/ Accepted 8 July 2004

Transcription of the yclJK operon, which encodes a potential two-component regulatory system, is activated in response to oxygen limitation in Bacillus subtilis. Northern blot analysis and assays of yclJ-lacZ reporter gene fusion activity revealed that the anaerobic induction is dependent on another two-component signal transduction system encoded by resDE. ResDE was previously shown to be required for the induction of anaerobic energy metabolism. Electrophoretic mobility shift assays and DNase I footprinting experiments showed that the response regulator ResD binds specifically to the yclJK regulatory region upstream of the transcriptional start site. In vitro transcription experiments demonstrated that ResD is sufficient to activate yclJ transcription. The phosphorylation of ResD by its sensor kinase, ResE, highly stimulates its activity as a transcriptional activator. Multiple nucleotide substitutions in the ResD binding regions of the yclJ promoter abolished ResD binding in vitro and prevented the anaerobic induction of yclJK in vivo. A weight matrix for the ResD binding site was defined by a bioinformatic approach. The results obtained suggest the existence of a new branch of the complex regulatory system employed for the adaptation of B. subtilis to anaerobic growth conditions.

* Corresponding author. Mailing address: Institute of Microbiology, 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.

{dagger} E.H. and H.G. contributed equally to this work.

Journal of Bacteriology, October 2004, p. 6477-6484, Vol. 186, No. 19
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.19.6477-6484.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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