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Applied and Environmental Microbiology, October 2005, p. 6282-6291, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6282-6291.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Genetic and Transcriptional Organization of the clpC Locus in Bifidobacterium breve UCC 2003

Marco Ventura,* Gerald F. Fitzgerald, and Douwe van Sinderen

Alimentary Pharmabiotic Centre and Department of Microbiology, Bioscience Institute, National University of Ireland, Western Road, Cork, Ireland

Received 7 December 2004/ Accepted 13 April 2005

A homolog of the clpC ATPase gene was identified in the genome of Bifidobacterium breve UCC 2003. Since this gene is very well conserved among eubacteria, we employed a PCR-based approach using primers based on highly conserved regions of ClpC proteins in order to identify homologous genes in other bifidobacterial species. Analysis by slot blot, Northern blot, and primer extension experiments showed that transcription of clpC is induced in response to moderate heat shock regimes. Moreover, we identified in the genome sequence of B. breve UCC 2003 a gene, designated clgR, which is predicted to encode a transcriptional regulator involved in regulation of the bifidobacterial clpC gene. The role of this protein in the regulation of B. breve UCC 2003 clpC gene expression was investigated by performing gel retardation experiments. We show that a biologically active ClgR molecule requires one or more proteinaceous coactivators to assist in the specific binding of ClgR to the clpC promoter region.

* Corresponding author. Mailing address: Department of Microbiology, National University of Ireland, Western Road, Cork, Ireland. Phone: 353-21-4901366. Fax: 353-21-4903031. E-mail: m.ventura{at}ucc.ie.

Applied and Environmental Microbiology, October 2005, p. 6282-6291, Vol. 71, No. 10
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.10.6282-6291.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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