J Korean Acad Oral Health.  2012 Dec;36(4):251-265.

Gene expression profile analysis of xylitol-sensitive and xylitol-resistant Streptococcus mutans in 0.5% glucose containing TYE media using DNA chip

Affiliations
  • 1Department of Preventive Dentistry, Kyungpook National University School of Dentistry, Daegu, Korea. kbsong@knu.ac.kr
  • 2Department of Dental Hygiene, Daegu Health College, Daegu, Korea.

Abstract


OBJECTIVES
Streptococcus mutans (S. mutans) is the major causative bacteria in dental caries. Xylitol is an effective anticarious natural sugar substitute by inhibiting the virulence of S. mutans. However, long-term xylitol consumption leads to the emergence of the xylitol-resistant S. mutans (XR). The aim of this study is to analyze the difference of gene expression profile of xylitol-sensitive S. mutans (XS) and XR in 0.5% glucose containing TYE media, using a DNA chip.
METHODS
S. mutans KCTC3065 was maintained in 0.5% glucose and 1% xylitol containing TYE media, during 30 days at 37degrees C 10% CO2 to form XR. The same procedures without xylitol were repeated for the formation of XS. Both XS and XR were cultured in 0.5% glucose with or without 1% xylitol containing TYE media overnight and total RNA was extracted. RNA from XS was labeled with Cy-3 dye as control, and XR were labeled with Cy-5 as references. DNA chip was hybridized for 18-20 h at 42degrees C.
RESULTS
A total of 277 genes of DNA chip data were significantly increased or decreased in XR. There is a total of 174 XR up-regulated genes in 0.5% glucose and 1% xylitol containing TYE media, and a total of 103 down-regulated genes. For compare with results of DNA chip, 11 in up-regulated genes and 10 in down-regulated were verified by RT-PCR. The most abundant increased genes in XR were related to cell envelope, cellular processes, DNA metabolism, transcription, and protein folding and stabilization. The decreased genes in XR were related to amino acid biosynthesis, toxin production and resistance, energy metabolism, ribosomal proteins synthesis, and signal transduction.
CONCLUSIONS
These results indicate that the difference of gene expression profile of XS and XR may be in existence. In particular, results of this study for XR up-regulated genes have a lot of similarities with the already published xylitol-related researches and other functional studies.

Keyword

DNA chip; Streptococcus mutans; Virulence; XR; XS; Xylitol

MeSH Terms

Bacteria
Chimera
Dental Caries
DNA
Energy Metabolism
Gene Expression
Glucose
Oligonucleotide Array Sequence Analysis
Protein Folding
Ribosomal Proteins
RNA
Streptococcus
Streptococcus mutans
Sweetening Agents
Transcriptome
Xylitol
DNA
Glucose
RNA
Ribosomal Proteins
Sweetening Agents
Xylitol

Figure

  • Fig. 1 The schema for XS and XR in 0.5% glucose and with or without 1% xylitol containing TYE media using DNA chip.

  • Fig. 2 A representative results of DNA chip for XS and XR in 0.5% glucose and with or without 1% xylitol containing TYE media. RNA samples from XS were labeled with Cy3 (green), and XR were labeled with Cy5 (red). DNA chip experiments were carried out two times. (A) 0704 experiment, (B) 0805 experiment.


Reference

1. Hamada S, Slade HD. Biology, immunology, and cariogenicity of Streptococcus mutans. Microbiol Rev. 1980. 44:331–384.
Article
2. Kuramitsu HK. Virulence factors of mutans streptococci: role of molecular genetics. Crit Rev Oral Biol Med. 1993. 4:159–176.
Article
3. Shemesh M, Tam A, Feldman M, Steinberg D. Differential expression profiles of Streptococcus mutans ftf, gtf and vicR genes in the presence of dietary carbohydrates at early and late exponential growth phases. Carbohydr Res. 2006. 341:2090–2097.
Article
4. Janda WM, Kuramitsu HK. Production of extracellular and cell-associated glucosyltransferase activity by Streptococcus mutans during growth on various carbon sources. Infect Immun. 1978. 19:116–122.
Article
5. Trahan L. Xylitol: a review of its action on mutans streptococci and dental plaque--its clinical significance. Int Dent J. 1995. 45:1 Suppl 1. S77–S92.
6. Holgerson PL, Sjöström I, Stecksén-Blicks C, Twetman S. Dental plaque formation and salivary mutans streptococci in schoolchildren after use of xylitol-containing chewing gum. Int J Paediatr Dent. 2007. 17:79–85.
Article
7. Han SK, Choi YH, Son EY, Song KB, Kim YJ, Nam SH. Prevention of dental caries by xylitol gum in pre-school children during 12-months. J Korean Acad Pediatr Dent. 2004. 31:159–168.
8. Trahan L, Bareil M, Gauthier L, Vadeboncoeur C. Transport and phosphorylation of xylitol by a fructose phosphotransferase system in Streptococcus mutans. Caries Res. 1985. 19:53–63.
Article
9. Lee YE, Choi YH, Jeong SH, Kim HS, Lee SH, Song KB. Morphological changes in Streptococcus mutans after chewing gum containing xylitol for twelve months. Curr Microbiol. 2009. 58:332–337.
Article
10. Trahan L, Mouton C. Selection for Streptococcus mutans with an altered xylitol transport capacity in chronic xylitol consumers. J Dent Res. 1987. 66:982–988.
Article
11. Trahan L, Söderling E, Dréan MF, Chevrier MC, Isokangas P. Effect of xylitol consumption on the plaque-saliva distribution of mutans streptococci and the occurrence and long-term survival of xylitol-resistant strains. J Dent Res. 1992. 71:1785–1791.
Article
12. Kim CC, Lee MN, Kim YJ, Lee SH. Quantitative comparison of mRNA expression of glucosyltransferase (GTF) between xylitol-resistant (XR) and xylitol-sensitive (XS) mutans streptococci. J Korean Acad Pediatr Dent. 2006. 33:77–84.
13. Lee HM, Kim JW, Jang KT, Lee SH, Hahn SH, Kim CC. A study on the cell property of xylitol-resistant Streptococcus mutans and xylitol-sensitive Streptococcus mutans. J Korean Acad Pediatr Dent. 2003. 30:554–562.
14. Assev S, Stig S, Scheie AA. Cariogenic traits in xylitol-resistant and xylitol-sensitive mutans streptococci. Oral Microbiol Immunol. 2002. 17:95–99.
Article
15. Cummings CA, Relman DA. Using DNA microarrays to study host-microbe interactions. Emerg Infect Dis. 2000. 6:513–525.
Article
16. Chen PM, Chen YY, Yu SL, Sher S, Lai CH, Chia JS. Role of GlnR in acid-mediated repression of genes encoding proteins involved in glutamine and glutamate metabolism in Streptococcus mutans. Appl Environ Microbiol. 2010. 76:2478–2486.
Article
17. Motegi M, Takagi Y, Yonezawa H, Hanada N, Terajima J, Watanabe H, et al. Assessment of genes associated with Streptococcus mutans biofilm morphology. Appl Environ Microbiol. 2006. 72:6277–6287.
Article
18. Shemesh M, Tam A, Steinberg D. Differential gene expression profiling of Streptococcus mutans cultured under biofilm and planktonic conditions. Microbiology. 2007. 153:1307–1317.
Article
19. Biswas I, Drake L, Biswas S. Regulation of gbpC expression in Streptococcus mutans. J Bacteriol. 2007. 189:6521–6531.
Article
20. Lynch DJ, Fountain TL, Mazurkiewicz JE, Banas JA. Glucan-binding proteins are essential for shaping Streptococcus mutans biofilm architecture. FEMS Microbiol Lett. 2007. 268:158–165.
Article
21. Sato Y, Yamamoto Y, Kizaki H. Xylitol-induced elevated expression of the gbpC gene in a population of Streptococcus mutans cells. Eur J Oral Sci. 2000. 108:538–545.
Article
22. Im SU, An SY, Choi YH, Song KB. Comparison of mRNA expression of gtf genes and adhesives ability of xylitol-sensitive and -resistant Streptococcus mutans by xylitol-treated concentrations. J Korean Acad Oral Health. 2012. 36:91–96.
23. Inagaki S, Matsumoto-Nakano M, Fujita K, Nagayama K, Funao J, Ooshima T. Effects of recombinase A deficiency on biofilm formation by Streptococcus mutans. Oral Microbiol Immunol. 2009. 24:104–108.
Article
24. Banas JA, Biswas S, Zhu M. Effects of DNA methylation on expression of virulence genes in Streptococcus mutans. Appl Environ Microbiol. 2011. 77:7236–7242.
Article
25. Shiroza T, Ueda S, Kuramitsu HK. Sequence analysis of the gtfB gene from Streptococcus mutans. J Bacteriol. 1987. 169:4263–4270.
Article
26. Vacca-Smith AM, Bowen WH. Binding properties of streptococcal glucosyltransferases for hydroxyapatite, saliva-coated hydroxyapatite, and bacterial surfaces. Arch Oral Biol. 1998. 43:103–110.
Article
27. Lee SH, Choi BK, Kim YJ. The cariogenic characters of xylitol-resistant and xylitol-sensitive Streptococcus mutans in biofilm formation with salivary bacteria. Arch Oral Biol. 2012. 57:697–703.
Article
28. Sato Y, Yamamoto Y, Kizaki H, Kuramitsu HK. Isolation, characterization and sequence analysis of the scrK gene encoding fructokinase of Streptococcus mutans. J Gen Microbiol. 1993. 139:921–927.
Article
29. Ramos JL, Martinez-Bueno M, Molina-Henares AJ, Terán W, Watanabe K, Zhang X, et al. The TetR family of transcriptional repressors. Microbiol Mol Biol Rev. 2005. 69:326–356.
Article
30. Jayaraman GC, Penders JE, Burne RA. Transcriptional analysis of the Streptococcus mutans hrcA, grpE and dnaK genes and regulation of expression in response to heat shock and environmental acidification. Mol Microbiol. 1997. 25:329–341.
Article
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