This application will investigate the potential pathogenic properties of Treponema denticola utilizing biochemical and molecular genetic approaches. Specifically, the role of proteases and a fibronectin-binding adhesin elaborated by these organisms in attachment and destruction of oral cells and invasion of basement membranes will be examined. These spirochete properties could play a role in the development of periodontal diseases. The gene coding for the T. denticola protease exhibiting type IV collagenase activity has been recently isolated in this laboratory. Further characterization of the gene will be accomplished following nucleotide sequencing. T. denticola clone banks generated in E. coli will also be screened for other protease genes. Spirochete mutants defective in individual proteases will then be sought following insertional inactivation of the genes, and introduction into T. denticola by electroporation or by conventional mutagenesis. These mutants will be utilized to examine the role of the enzymes in model cellular attachment and invasion systems. Likewise, the gene coding for the fibronectin-binding adhesin (FBA) will be isolated in lambda gtll clone banks. The protein product will be purified and utilized to produce monospecific antibody to localize the adhesin in T. denticola. The gene will be sequenced and the deduced amino acid sequence utilized to identify functional domains of the protein following site-directed mutagenesis. In addition, the gene will be used to generate FBA-negative mutants of the spirochete for assessment of the role of the gene product in attachment to basement membranes. These approaches should provide new information regarding the pathogenic potential of T. denticola and its possible role in periodontal diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
7R01DE009821-03
Application #
3223545
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Project Start
1992-02-20
Project End
1997-02-19
Budget Start
1993-07-01
Budget End
1994-02-19
Support Year
3
Fiscal Year
1993
Total Cost
Indirect Cost
Name
State University of New York at Buffalo
Department
Type
Schools of Dentistry
DUNS #
038633251
City
Buffalo
State
NY
Country
United States
Zip Code
14260
He, X; Lux, R; Kuramitsu, H K et al. (2009) Achieving probiotic effects via modulating oral microbial ecology. Adv Dent Res 21:53-6
Cameron, Caroline E; Kuroiwa, Janelle M Y; Yamada, Mitsunori et al. (2008) Heterologous expression of the Treponema pallidum laminin-binding adhesin Tp0751 in the culturable spirochete Treponema phagedenis. J Bacteriol 190:2565-71
Kuramitsu, Howard K; He, Xuesong; Lux, Renate et al. (2007) Interspecies interactions within oral microbial communities. Microbiol Mol Biol Rev 71:653-70
Han, Yiping W; Ikegami, Akihiko; Rajanna, Chythanya et al. (2005) Identification and characterization of a novel adhesin unique to oral fusobacteria. J Bacteriol 187:5330-40
Yamada, Mitsunori; Ikegami, Akihiko; Kuramitsu, Howard K (2005) Synergistic biofilm formation by Treponema denticola and Porphyromonas gingivalis. FEMS Microbiol Lett 250:271-7
Kuramitsu, Howard K; Chen, Wen; Ikegami, Aki (2005) Biofilm formation by the periodontopathic bacteria Treponema denticola and Porphyromonas gingivalis. J Periodontol 76:2047-51
Ishihara, Kazuyuki; Kuramitsu, Howard K; Okuda, Katsuji (2004) A 43-kDa protein of Treponema denticola is essential for dentilisin activity. FEMS Microbiol Lett 232:181-8
Ikegami, Akihiko; Honma, Kiyonobu; Sharma, Ashu et al. (2004) Multiple functions of the leucine-rich repeat protein LrrA of Treponema denticola. Infect Immun 72:4619-27
Chauhan, Sarita; Kuramitsu, Howard K (2004) Sequence analysis of plasmid pTS1 isolated from oral spirochetes. Plasmid 51:61-5
Vesey, Peter M; Kuramitsu, Howard K (2004) Genetic analysis of Treponema denticola ATCC 35405 biofilm formation. Microbiology 150:2401-7

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