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Journal of Bacteriology, April 2009, p. 2362-2370, Vol. 191, No. 7
0021-9193/09/$08.00+0     doi:10.1128/JB.01616-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Copper Acquisition Is Mediated by YcnJ and Regulated by YcnK and CsoR in Bacillus subtilis{triangledown}

Shashi Chillappagari,1 Marcus Miethke,1,2 Hein Trip,3 Oscar P. Kuipers,3 and Mohamed A. Marahiel1*

Fachbereich Chemie/Biochemie der Philipps-Universität Marburg, Hans-Meerwein-Str., D-35032 Marburg, Germany,1 Lehrstuhl für Biologische Chemie, Technische Universität München, Freising-Weihenstephan, Germany,2 Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands3

Received 14 November 2008/ Accepted 9 January 2009

Copper is an essential cofactor for many enzymes, and at over a threshold level, it is toxic for all organisms. To understand the mechanisms underlying copper homeostasis of the gram-positive bacterium Bacillus subtilis, we have performed microarray studies under copper-limiting conditions. These studies revealed that the ycnJ gene encodes a protein that plays an important role in copper metabolism, as it shows a significant, eightfold upregulation under copper-limiting conditions and its disruption causes a growth-defective phenotype under copper deprivation as well as a reduced intracellular content of copper. Native gel shift experiments with the periplasmic N-terminal domain of the YcnJ membrane protein (135 residues) disclosed its strong affinity to Cu(II) ions in vitro. Inspection of the upstream sequence of ycnJ revealed that the ycnK gene encodes a putative transcriptional regulator, whose deletion caused an elevated expression of ycnJ, especially under conditions of copper excess. Further studies demonstrated that the recently identified copper efflux regulator CsoR also is involved in the regulation of ycnJ expression, leading to a new model for copper homeostasis in B. subtilis.

* Corresponding author. Mailing address: Fachbereich Chemie/Biochemie der Philipps-Universität Marburg, Hans-Meerwein-Str., D-35032 Marburg, Germany. Phone: 49 6421 2825722. Fax: 49 6421 2822191. E-mail: marahiel{at}staff.uni-marburg.de

{triangledown} Published ahead of print on 23 January 2009.

Journal of Bacteriology, April 2009, p. 2362-2370, Vol. 191, No. 7
0021-9193/09/$08.00+0     doi:10.1128/JB.01616-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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