Poster Presentation

Poster Presentation

PR100 - Small Non-Coding RNAs Functionally Contribute to the Negative Regulation of Type IV Pilus Retraction ATPase PilT and Biofilm Formation in Streptococcus Sanguinis

Thursday, April 26
10:00 AM - 1:00 PM
Room: Exhibit Hall

Objective: Streptococcus sanguinis is a commensal pioneer colonizer in oral biofilm and is also implicated in endodontic infections. However, the mechanisms of adaptation of S. sanguinis to the root canal environment are poorly defined. The adaptive response to environmental change can be transduced by the CiaRH two-component regulatory system in streptococcal species. Recently, cia-dependent small RNAs (csRNAs), a group of non-coding RNAs controlled by CiaRH system, were identified and their roles in bacterial adaptation to novel environments have been received much attention. In this study, we performed functional analyses of csRNAs in S. sanguinis. Methods: S. sanguinis mutants were created by homologous recombination. Gene expression was measured by real-time RT-PCR, and reporter gene activity was assessed by a luciferase reporter assay. Interaction of RNA molecules was examined by electrophoretic mobility shift assay (EMSA). Biofilm-forming ability was evaluated using 1% sucrose-containing medium. Results: Using the TargetRNA2 computational search tool, pilT,a type IV pilus retraction ATPase gene, was identified as the most probable target for csRNA1-1 and csRNA1-2. A significant increase in expression of pilT in the csRNA1-1, csRNA1-2 double deletion mutant was observed with real-time RT-PCR analysis and luciferase assay. The EMSA results showed direct binding of csRNA1-1 with pilT mRNA. Biofilm-forming ability of csRNA1-1, csRNA1-2 double deletion mutant increased significantly compared to wild-type strain. Conclusion: These results suggest that csRNA1-1 and csRNA1-2 negatively regulate the expression of pilT mRNA via base-pairing, and may contribute to the regulation of virulence properties, including adaptation to the endodontic environment, in S. sanguinis

Chiaki Ota

PhD Student with no professional degree

Chiaki Ota, D.D.S, is Ph. D. candidate in Showa University. I received my Doctor of Dental Surgery degree from Showa University in 2013. My main research project is "biomolecular and functional analysis of non-coding RNA in bacteria".

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Noriyuki Suzuki

Associate professor

Noriyuki Suzuki, D.D.S, Ph. D. He received his Doctor of Dental Surgery degree from Tokyo Medical and Dental University and his Ph.D degree from Tokyo Medical and Dental University. He is currently serve as an accosiate professor, Department of Conservative Dentistry, Division of Endodontology in Showa University School of Dentistry.

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Takashi Miyazaki

Professor

Takashi Miyazaki, D.D.S, Ph. D. He received his Doctor of Dental Surgery degree from Tokyo Medical and Dental University and his Ph.D degree from Tokyo Medical and Dental University. He is currently serve as a professor, Division of Biomaterials and Engineering, Department of Conservative Dentistry, Showa University School of Dentistry.

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PR100 - Small Non-Coding RNAs Functionally Contribute to the Negative Regulation of Type IV Pilus Retraction ATPase PilT and Biofilm Formation in Streptococcus Sanguinis



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