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Microbiology 153 (2007), 866-876; DOI  10.1099/mic.0.29275-0
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Microbiology 153 (2007), 866-876; DOI  10.1099/mic.0.29275-0
© 2007 Society for General Microbiology

Loss of adherence ability to human gingival epithelial cells in S-layer protein-deficient mutants of Tannerella forsythensis

Junpei Sakakibara1,2, Keiji Nagano1,{dagger}, Yukitaka Murakami1, Naoya Higuchi3, Hiroshi Nakamura3, Kazuo Shimozato2 and Fuminobu Yoshimura1

1 Department of Microbiology, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan
2 Department of Oral and Maxillofacial Surgery II, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan
3 Department of Endodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan

Correspondence
Yukitaka Murakami
yukimu{at}dpc.agu.ac.jp

Tannerella forsythensis, one of the important pathogens in periodontal disease, has a typical surface layer (S-layer) consisting of regularly arrayed subunits outside the outer membrane. The S-layer in T. forsythensis is suggested to be associated with haemagglutinating activity, adhesion and invasion of host cells; however, its precise functions have been unknown. ORFs encoding the major S-layer proteins (230 and 270 kDa) of T. forsythensis ATCC 43037, tfsA and tfsB, respectively, following the names in a recent report [Lee, S.-W., Sabet, M., Um, H. S., Yang, L., Kim, H. C. & Zhu, W. (2006). Gene 371, 102–111] were determined. To verify the function of the S-layer proteins, three mutants with tfsA, tfsB, or both deleted were successfully constructed by a PCR-based overlapping method. S-layer proteins were completely lost in the double mutant. The single-deletion mutants appeared to lose one of the 230 and 270 kDa proteins. Thin-section microscopy clearly revealed that the 230 and 270 kDa proteins composed the S-layer. Although the S-layer proteins may be weakly related to haemagglutinating activity, these proteins were highly responsible for adherence to human gingival epithelial cells (Ca9-22) and KB cells. These results suggest that the S-layer proteins in T. forsythensis play an important role in the initiation stage of oral infection including periodontal disease.


Abbreviations: CAT, chloramphenicol acetyltransferase; CBB, Coomassie brilliant blue; Cp, chloramphenicol; TIGR, The Institute for Genomic Research; TRITC, tetramethyl rhodamine isothiocyanate

{dagger}Present address: Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3202, USA.




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