This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Scragg, M. A.
Right arrow Articles by Curtis, M. A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Scragg, M. A.
Right arrow Articles by Curtis, M. A.

 Previous Article  |  Next Article 

Infection and Immunity, October 2002, p. 5740-5750, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5740-5750.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Nuclear Targeting of Porphyromonas gingivalis W50 Protease in Epithelial Cells

Margaret A. Scragg,1* Asil Alsam,1 Minnie Rangarajan,2 Jennifer M. Slaney,2 Philip Shepherd,3 David M. Williams,1 and Michael A. Curtis2

Department of Clinical and Diagnostic Oral Sciences (Oral Pathology),1 MRC Molecular Pathogenesis Group, Department of Medical Microbiology, Barts and The London School of Medicine and Dentistry, Queen Mary, University of London, London E1 2AD,2 Department of Immunology, United Medical and Dental Schools, London SE1 9RT, United Kingdom3

Received 7 February 2002/ Returned for modification 26 March 2002/ Accepted 24 June 2002

Porphyromonas gingivalis is an important pathogen associated with destructive periodontal disease and is able to invade the epithelial cell barrier. Its cysteine proteases are recognized as major virulence factors, and in this study, we examined the interaction of the arginine-specific protease with epithelial cells in culture. Three cell lines (KB, HeLa, and SCC4) were incubated with strain W50 culture supernatant; stained with monoclonal antibody 1A1, which recognizes an epitope on the adhesin (ß) component of the cysteine protease-adhesin ({alpha}/ß) heterodimer; and viewed using immunofluorescence microscopy. Within 1 h, the protease traversed the plasma membrane and was localized around the nucleus before becoming concentrated in the cytoplasm after 24 to 48 h. In contrast, the purified arginine-specific heterodimeric protease (HRgpA) rapidly entered the nucleus within 15 to 30 min. This nuclear targeting (i) was seen with active and N{alpha}-p-tosyl-L-lysine chloromethyl ketone (TLCK)-inactivated HRgpA, indicating it was independent of the proteolytic activity; (ii) occurred at both 4 and 37°C; and (iii) failed to occur with the monomeric protease (RgpAcat), indicating the importance of the adhesin chain of the HRgpA protease to this process. Rapid cell entry was also observed with recombinant catalytic ({alpha}) and adhesin (ß) chains, with the latter again targeting the nuclear area. After 48 h of incubation with HRgpA, significant dose-dependent stimulation of metabolic activity was observed (measured by reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide), and a doubling of mitotic activity combined with the presence of apoptotic cells indicated that HRgpA may interfere with cell cycle control mechanisms. These effects were seen with both active and TLCK-inactivated protease, confirming that they were not dependent on proteolytic activity, and thus provide new insights into the functioning of this P. gingivalis protease.

* Corresponding author. Mailing address: Department of Clinical and Diagnostic Oral Sciences (Oral Pathology), Barts and The London School of Medicine and Dentistry, Turner St., London E1 2AD, United Kingdom. Phone: 44(0) 20 7882 7130 or 44(0) 20 7882 7154. Fax: 44(0) 20 7882 7153. E-mail: m.a.scragg{at}qmul.ac.uk.

Editor: D. L. Burns

Infection and Immunity, October 2002, p. 5740-5750, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5740-5750.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Furuta, N., Takeuchi, H., Amano, A. (2009). Entry of Porphyromonas gingivalis Outer Membrane Vesicles into Epithelial Cells Causes Cellular Functional Impairment. Infect. Immun. 77: 4761-4770 [Abstract] [Full Text]  
  • Furuta, N., Tsuda, K., Omori, H., Yoshimori, T., Yoshimura, F., Amano, A. (2009). Porphyromonas gingivalis Outer Membrane Vesicles Enter Human Epithelial Cells via an Endocytic Pathway and Are Sorted to Lysosomal Compartments. Infect. Immun. 77: 4187-4196 [Abstract] [Full Text]  
  • Andrian, E., Grenier, D., Rouabhia, M. (2006). Porphyromonas gingivalis-Epithelial Cell Interactions in Periodontitis. JDR 85: 392-403 [Abstract] [Full Text]  
  • Yilmaz, O., Verbeke, P., Lamont, R. J., Ojcius, D. M. (2006). Intercellular Spreading of Porphyromonas gingivalis Infection in Primary Gingival Epithelial Cells. Infect. Immun. 74: 703-710 [Abstract] [Full Text]  
  • Hall, L. M. C., Fawell, S. C., Shi, X., Faray-Kele, M.-C., Aduse-Opoku, J., Whiley, R. A., Curtis, M. A. (2005). Sequence Diversity and Antigenic Variation at the rag Locus of Porphyromonas gingivalis. Infect. Immun. 73: 4253-4262 [Abstract] [Full Text]  
  • Yun, P. L. W., Decarlo, A. A., Chapple, C. C., Hunter, N. (2005). Functional Implication of the Hydrolysis of Platelet Endothelial Cell Adhesion Molecule 1 (CD31) by Gingipains of Porphyromonas gingivalis for the Pathology of Periodontal Disease. Infect. Immun. 73: 1386-1398 [Abstract] [Full Text]  
  • Sheets, S. M., Potempa, J., Travis, J., Casiano, C. A., Fletcher, H. M. (2005). Gingipains from Porphyromonas gingivalis W83 Induce Cell Adhesion Molecule Cleavage and Apoptosis in Endothelial Cells. Infect. Immun. 73: 1543-1552 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»