1887

Abstract

Potassium (K) is the most abundant cation in dental plaque fluid. Previously, we reported the link between K transport via Trk2 in Streptococcus mutans and its two critical virulence attributes: acid tolerance and surface adhesion. Herein, we build further on the intimate link between K levels and S. mutans biology. High (>25 mM) versus low (≤5 mM) K concentrations in the growth medium affected conformational epitopes of cell surface-localized adhesin P1. At low K, the expression of stress response elements gcrR and codY, cell-adhesion-associated genes such as spaP and metabolism-associated genes such as bglP was induced at stationary phase (P<0.05), suggesting that K-mediated regulation is growth phase-dependent and stress-sensitive. Production of the newly discovered secretory protein encoded by SMU_63 c was strongly dependent on the availability of K and growth phase. This protein is a newly discovered regulator of genetic competence and biofilm cell density. Thus, the influence of K on DNA transformation efficiency was also examined. Compared with 25 mM K concentration, the presence of low K reduced the transformation frequency by 100-fold. Genetic transformation was abolished in a strain lacking a Trk2 system under all K concentrations tested. Consistent with these findings, repression of competence-associated genes, comS and comX, was observed under low environmental K conditions and in the strain lacking Trk2. Taken together, these results highlight a pivotal role for environmental K as a regulatory cation that modulates stress responses and genetic transformation in S. mutans.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000458
2017-06-22
2019-10-21
Loading full text...

Full text loading...

/deliver/fulltext/micro/163/5/719.html?itemId=/content/journal/micro/10.1099/mic.0.000458&mimeType=html&fmt=ahah

References

  1. Lemos JA, Burne RA. A model of efficiency: stress tolerance by Streptococcus mutans. Microbiology 2008;154:3247–3255 [CrossRef][PubMed]
    [Google Scholar]
  2. Li YH, Lau PC, Lee JH, Ellen RP, Cvitkovitch DG. Natural genetic transformation of Streptococcus mutans growing in biofilms. J Bacteriol 2001;183:897–908 [CrossRef][PubMed]
    [Google Scholar]
  3. Prudhomme M, Attaiech L, Sanchez G, Martin B, Claverys JP. Antibiotic stress induces genetic transformability in the human pathogen Streptococcus pneumoniae. Science 2006;313:89–92 [CrossRef][PubMed]
    [Google Scholar]
  4. Mashburn-Warren L, Morrison DA, Federle MJ. A novel double-tryptophan peptide pheromone controls competence in Streptococcus spp. via an Rgg regulator. Mol Microbiol 2010;78:589–606 [CrossRef][PubMed]
    [Google Scholar]
  5. Fontaine L, Goffin P, Dubout H, Delplace B, Baulard A et al. Mechanism of competence activation by the ComRS signalling system in streptococci. Mol Microbiol 2013;87:1113–1132 [CrossRef][PubMed]
    [Google Scholar]
  6. Senadheera D, Cvitkovitch DG. Quorum sensing and biofilm formation by Streptococcus mutans. Adv Exp Med Biol 2008;631:178–188 [CrossRef][PubMed]
    [Google Scholar]
  7. Reck M, Tomasch J, Wagner-Döbler I. The alternative sigma factor sigX controls bacteriocin synthesis and competence, the two quorum sensing regulated traits in Streptococcus mutans. PLoS Genet 2015;11:e1005353 [CrossRef][PubMed]
    [Google Scholar]
  8. Cowman RA, Fitzgerald RJ. Potassium requirement of oral streptococci. J Dent Res 1976;55:709 [CrossRef][PubMed]
    [Google Scholar]
  9. Moreno EC, Margolis HC. Composition of human plaque fluid. J Dent Res 1988;67:1181–1189 [CrossRef][PubMed]
    [Google Scholar]
  10. Tatevossian A, Gould CT. The composition of the aqueous phase in human dental plaque. Arch Oral Biol 1976;21:319–323 [CrossRef][PubMed]
    [Google Scholar]
  11. Binepal G, Gill K, Crowley P, Cordova M, Brady LJ et al. Trk2 potassium transport system in Streptococcus mutans and its role in potassium homeostasis, biofilm formation, and stress tolerance. J Bacteriol 2016;198:1087–1100 [CrossRef][PubMed]
    [Google Scholar]
  12. van Nieuwenhoven MH, Hellingwerf KJ, Venema G, Konings WN. Role of proton motive force in genetic transformation of Bacillus subtilis. J Bacteriol 1982;151:771–776[PubMed]
    [Google Scholar]
  13. Kohoutova M. Kinetics of the potassium- and sodium-activated infection of a transforming deoxyribonucleic acid in pneumococcus. J Gen Microbiol 1965;38:211–219 [CrossRef][PubMed]
    [Google Scholar]
  14. Fujiwara S, Kobayashi S, Nakayama H. Development of a minimal medium for Streptococcus mutans. Arch Oral Biol 1978;23:601–602 [CrossRef][PubMed]
    [Google Scholar]
  15. van de Rijn I, Kessler RE. Growth characteristics of group A streptococci in a new chemically defined medium. Infect Immun 1980;27:444–448[PubMed]
    [Google Scholar]
  16. Lau PC, Sung CK, Lee JH, Morrison DA, Cvitkovitch DG. PCR ligation mutagenesis in transformable streptococci: application and efficiency. J Microbiol Methods 2002;49:193–205 [CrossRef][PubMed]
    [Google Scholar]
  17. Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001;29:e45 [CrossRef][PubMed]
    [Google Scholar]
  18. Besingi RN, Wenderska IB, Senadheera DB, Cvitkovitch DG, Long JR et al. Functional amyloids in Streptococcus mutans, their use as targets of biofilm inhibition and initial characterization of SMU_63c. Microbiology 2017;163:488–501 [CrossRef][PubMed]
    [Google Scholar]
  19. Larson MR, Rajashankar KR, Patel MH, Robinette RA, Crowley PJ et al. Elongated fibrillar structure of a streptococcal adhesin assembled by the high-affinity association of alpha- and PPII-helices. Proc Natl Acad Sci USA 2010;107:5983–5988 [CrossRef][PubMed]
    [Google Scholar]
  20. Heim KP, Crowley PJ, Brady LJ. An intramolecular interaction involving the N terminus of a streptococcal adhesin affects its conformation and adhesive function. J Biol Chem 2013;288:13762–13774 [CrossRef][PubMed]
    [Google Scholar]
  21. Heim KP, Crowley PJ, Long JR, Kailasan S, Mckenna R et al. An intramolecular lock facilitates folding and stabilizes the tertiary structure of Streptococcus mutans adhesin P1. Proc Natl Acad Sci USA 2014;111:15746–15751 [CrossRef][PubMed]
    [Google Scholar]
  22. Palmer SR, Crowley PJ, Oli MW, Ruelf MA, Michalek SM et al. YidC1 and YidC2 are functionally distinct proteins involved in protein secretion, biofilm formation and cariogenicity of Streptococcus mutans. Microbiology 2012;158:1702–1712 [CrossRef][PubMed]
    [Google Scholar]
  23. Son M, Shields RC, Ahn SJ, Burne RA, Hagen SJ. Bidirectional signaling in the competence regulatory pathway of Streptococcus mutans. FEMS Microbiol Lett 2015;362:fnv159 [CrossRef][PubMed]
    [Google Scholar]
  24. Loesche WJ. Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50:353–380[PubMed]
    [Google Scholar]
  25. Bowen WH. Dental caries - not just holes in teeth! A perspective. Mol Oral Microbiol 2016;31:228–233 [CrossRef][PubMed]
    [Google Scholar]
  26. Ajdić D, Pham VT. Global transcriptional analysis of Streptococcus mutans sugar transporters using microarrays. J Bacteriol 2007;189:5049–5059 [CrossRef][PubMed]
    [Google Scholar]
  27. Bettenbrock K, Sauter T, Jahreis K, Kremling A, Lengeler JW et al. Correlation between growth rates, EIIACrr phosphorylation, and intracellular cyclic AMP levels in Escherichia coli K-12. J Bacteriol 2007;189:6891–6900 [CrossRef][PubMed]
    [Google Scholar]
  28. Hogema BM, Arents JC, Bader R, Eijkemans K, Yoshida H et al. Inducer exclusion in Escherichia coli by non-PTS substrates: the role of the PEP to pyruvate ratio in determining the phosphorylation state of enzyme IIAGlc. Mol Microbiol 1998;30:487–498 [CrossRef][PubMed]
    [Google Scholar]
  29. Lee CR, Cho SH, Yoon MJ, Peterkofsky A, Seok YJ. Escherichia coli enzyme IIANtr regulates the K+ transporter TrkA. Proc Natl Acad Sci USA 2007;104:4124–4129 [CrossRef][PubMed]
    [Google Scholar]
  30. Lüttmann D, Heermann R, Zimmer B, Hillmann A, Rampp IS et al. Stimulation of the potassium sensor KdpD kinase activity by interaction with the phosphotransferase protein IIA(Ntr) in Escherichia coli. Mol Microbiol 2009;72:978–994 [CrossRef][PubMed]
    [Google Scholar]
  31. Bahr T, Lüttmann D, März W, Rak B, Görke B. Insight into bacterial phosphotransferase system-mediated signaling by interspecies transplantation of a transcriptional regulator. J Bacteriol 2011;193:2013–2026 [CrossRef][PubMed]
    [Google Scholar]
  32. Lemos JA, Abranches J, Burne RA. Responses of cariogenic streptococci to environmental stresses. Curr Issues Mol Biol 2005;7:95–107[PubMed]
    [Google Scholar]
  33. Lemos JA, Lin VK, Nascimento MM, Abranches J, Burne RA. Three gene products govern (p)ppGpp production by Streptococcus mutans. Mol Microbiol 2007;65:1568–1581 [CrossRef][PubMed]
    [Google Scholar]
  34. Senadheera MD, Guggenheim B, Spatafora GA, Huang YC, Choi J et al. A VicRK signal transduction system in Streptococcus mutans affects gtfBCD, gbpB, and ftf expression, biofilm formation, and genetic competence development. J Bacteriol 2005;187:4064–4076 [CrossRef][PubMed]
    [Google Scholar]
  35. Lemos JA, Nascimento MM, Lin VK, Abranches J, Burne RA. Global regulation by (p)ppGpp and CodY in Streptococcus mutans. J Bacteriol 2008;190:5291–5299 [CrossRef][PubMed]
    [Google Scholar]
  36. Sonenshein AL. CodY, a global regulator of stationary phase and virulence in Gram-positive bacteria. Curr Opin Microbiol 2005;8:203–207 [CrossRef][PubMed]
    [Google Scholar]
  37. Dashper SG, Reynolds EC. Branched-chain amino acid transport in Streptococcus mutans Ingbritt. Oral Microbiol Immunol 1993;8:167–171 [CrossRef][PubMed]
    [Google Scholar]
  38. Xing RY, Whitman WB. Characterization of enzymes of the branched-chain amino acid biosynthetic pathway in Methanococcus spp. J Bacteriol 1991;173:2086–2092 [CrossRef][PubMed]
    [Google Scholar]
  39. Banas JA, Vickerman MM. Glucan-binding proteins of the oral streptococci. Crit Rev Oral Biol Med 2003;14:89–99[PubMed][CrossRef]
    [Google Scholar]
  40. Bowen WH, Koo H. Biology of Streptococcus mutans-derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms. Caries Res 2011;45:69–86 [CrossRef][PubMed]
    [Google Scholar]
  41. Russell MW, Mansson-Rahemtulla B. Interaction between surface protein antigens of Streptococcus mutans and human salivary components. Oral Microbiol Immunol 1989;4:106–111 [CrossRef][PubMed]
    [Google Scholar]
  42. Novichkov PS, Kazakov AE, Ravcheev DA, Leyn SA, Kovaleva GY et al. RegPrecise 3.0-a resource for genome-scale exploration of transcriptional regulation in bacteria. BMC Genomics 2013;14:745 [CrossRef][PubMed]
    [Google Scholar]
  43. Dunny GM, Lee LN, Leblanc DJ. Improved electroporation and cloning vector system for gram-positive bacteria. Appl Environ Microbiol 1991;57:1194–1201[PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000458
Loading
/content/journal/micro/10.1099/mic.0.000458
Loading

Data & Media loading...

Supplements

Supplementary File 1

PDF
This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error