Loss of adherence ability to human gingival epithelial cells in S-layer protein-deficient mutants of Tannerella forsythensis.
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.
Sakakibara
Junpei
J
Department of Oral and Maxillofacial Surgery II, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.|Department of Microbiology, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Nagano
Keiji
K
Department of Microbiology, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Murakami
Yukitaka
Y
Department of Microbiology, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Higuchi
Naoya
N
Department of Endodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Nakamura
Hiroshi
H
Department of Endodontics, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Shimozato
Kazuo
K
Department of Oral and Maxillofacial Surgery II, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
Yoshimura
Fuminobu
F
Department of Microbiology, School of Dentistry, Aichi-Gakuin University, Nagoya, Aichi 464-8650, Japan.
D018829:Adhesins, Bacterial / Q000235:genetics / Q000502:physiology*; D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D041963:Bacteroidetes / Q000235:genetics* / Q000472:pathogenicity* / Q000648:ultrastructure; D015153:Blotting, Western; D004847:Epithelial Cells / Q000382:microbiology*; D017353:Gene Deletion; D005881:Gingiva / Q000166:cytology / Q000382:microbiology*; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D008970:Molecular Weight; D016133:Polymerase Chain Reaction / Q000379:methods; D037521:Virulence Factors / Q000235:genetics / Q000502:physiology
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D008562:Membrane Glycoproteins; C055764:S-layer proteins; D037521:Virulence Factors