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Journal of Bacteriology, February 2008, p. 1436-1446, Vol. 190, No. 4
0021-9193/08/$08.00+0     doi:10.1128/JB.01632-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Role of the Clp System in Stress Tolerance, Biofilm Formation, and Intracellular Invasion in Porphyromonas gingivalis

Cindy A. Capestany,¹ Gena D. Tribble,^1, Kazuhiko Maeda,¹ Donald R. Demuth,² and Richard J. Lamont^1*

Department of Oral Biology and Center for Molecular Microbiology, College of Dentistry, University of Florida, Gainesville, Florida 32610,¹ Department of Periodontics, Endodontics and Dental Hygiene, University of Louisville School of Dentistry, Louisville, Kentucky 40292²

Received 8 October 2007/ Accepted 23 November 2007

Clp proteases and chaperones are ubiquitous among prokaryotes and eukaryotes, and in many pathogenic bacteria the Clp stress response system is also involved in regulation of virulence properties. In this study, the roles of ClpB, ClpC, and ClpXP in stress resistance, homotypic and heterotypic biofilm formation, and intracellular invasion in the oral opportunistic pathogen Porphyromonas gingivalis were investigated. Absence of ClpC and ClpXP, but not ClpB, resulted in diminished tolerance to high temperatures. Response to oxidative stress was not affected by the loss of any of the Clp proteins. The clpC and clpXP mutants demonstrated elevated monospecies biofilm formation, and the absence of ClpXP also enhanced heterotypic P. gingivalis-Streptococcus gordonii biofilm formation. All clp mutants adhered to gingival epithelial cells to the same level as the wild type; however, ClpC and ClpXP were found to be necessary for entry into host epithelial cells. ClpB did not play a role in entry but was required for intracellular replication and survival. ClpXP negatively regulated the surface exposure of the minor fimbrial (Mfa) protein subunit of P. gingivalis, which stimulates biofilm formation but interferes with epithelial cell entry. Collectively, these results show that the Clp protease complex and chaperones control several processes that are important for the colonization and survival of P. gingivalis in the oral cavity.

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* Corresponding author. Mailing address: Department of Oral Biology, College of Dentistry, University of Florida, Gainesville, FL 32610-0424. Phone: (352) 392-5067. Fax: (352) 392-2361. E-mail: rlamont{at}dental.ufl.edu

Published ahead of print on 7 December 2007.

Present address: Department of Periodontics, Dental Branch, University of Texas Health Sciences Center at Houston, Houston, TX.

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Journal of Bacteriology, February 2008, p. 1436-1446, Vol. 190, No. 4
0021-9193/08/$08.00+0     doi:10.1128/JB.01632-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

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