Oral Biology pp 219-232 | Cite as

Natural Transformation of Oral Streptococci by Use of Synthetic Pheromones

  • Gabriela Salvadori
  • Roger Junges
  • Rabia Khan
  • Heidi A. Åmdal
  • Donald A. Morrison
  • Fernanda C. PetersenEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1537)


The discovery that Streptococcus pneumoniae uses a competence-stimulating peptide (CSP) to induce competence for natural transformation, and that other species of the mitis and the anginosus streptococcal groups use a similar system, has expanded the tools to explore gene function and regulatory pathways in streptococci. Two other classes of pheromones have been discovered since then, comprising the bacteriocin-inducing peptide class found in Streptococcus mutans (also named CSP, although different from the former) and the SigX-inducing peptides (XIP), in the mutans, salivarius, bovis, and pyogenes groups of streptococci. The three classes of peptide pheromones can be ordered from peptide synthesis services at affordable prices, and used in transformation assays to obtain competent cultures consistently at levels usually higher than those achieved during spontaneous competence. In this chapter, we present protocols for natural transformation of oral streptococci that are based on the use of synthetic pheromones, with examples of conditions optimized for transformation of S. mutans and Streptococcus mitis.

Key words

Oral streptococcus Streptococcus mutans Streptococcus mitis Competence Natural transformation Competence-stimulating peptide (CSP) SigX-inducing peptide (XIP) 



This work was partially supported by the National Science Foundation, grant no. MCB-1020863, by the Faculty of Dentistry, University of Oslo, and by the Norwegian surveillance system for antibiotic resistance in microbes (Norsk overvåkingssystem for antibiotikaresistens hos mikrober—NORM).


  1. 1.
    Federle MJ, Morrison DA (2012) One if by land, two if by sea: signalling to the ranks with CSP and XIP. Mol Microbiol 86:241–245CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Fontaine L, Wahl A, Flechard M, Mignolet J, Hols P (2015) Regulation of competence for natural transformation in streptococci. Infection, genetics and evolution. J Mol Epidemiol Evol Genet Infect Dis 33:343–360CrossRefGoogle Scholar
  3. 3.
    Johnsborg O, Eldholm V, Håvarstein LS (2007) Natural genetic transformation: prevalence, mechanisms and function. Res Microbiol 158:767–778CrossRefPubMedGoogle Scholar
  4. 4.
    Håvarstein LS, Hakenbeck R, Gaustad P (1997) Natural competence in the genus Streptococcus: evidence that streptococci can change pherotype by interspecies recombinational exchanges. J Bacteriol 179:6589–6594CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Morrison DA (1997) Streptococcal competence for genetic transformation: regulation by peptide pheromones. Microb Drug Resist 3:27–37CrossRefPubMedGoogle Scholar
  6. 6.
    Khan R, Rukke HV, Ricomini Filho AP, Fimland G, Arntzen MO, Thiede B, Petersen FC (2012) Extracellular identification of a processed type II ComR/ComS pheromone of Streptococcus mutans. J Bacteriol 194:3781–3788CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Mashburn-Warren L, Morrison DA, Federle MJ (2010) A novel double-tryptophan peptide pheromone controls competence in Streptococcus spp. via an Rgg regulator. Mol Microbiol 78589–78606Google Scholar
  8. 8.
    Fontaine L, Boutry C, de Frahan MH, Delplace B, Fremaux C, Horvath P, Boyaval P, Hols P (2010) A novel pheromone quorum-sensing system controls the development of natural competence in Streptococcus thermophilus and Streptococcus salivarius. J Bacteriol 192:1444–1454CrossRefPubMedGoogle Scholar
  9. 9.
    Monnet V, Juillard V, Gardan R (2016) Peptide conversations in Gram-positive bacteria. Crit Rev Microbiol 42:339–351PubMedGoogle Scholar
  10. 10.
    Lemme A, Grobe L, Reck M, Tomasch J, Wagner-Dobler I (2011) Subpopulation-Specific transcriptome analysis of competence stimulating peptide induced Streptococcus mutans. J Bacteriol 193:1863–1877CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Son M, Shields RC, Ahn SJ, Burne RA, Hagen SJ (2015) Bidirectional signaling in the competence regulatory pathway of Streptococcus mutans. FEMS Microbiol Lett 362. Doi:  10.1093/femsle/fnv159
  12. 12.
    Chen JD, Morrison DA (1987) Modulation of competence for genetic transformation in Streptococcus pneumoniae. J Gen Microbiol 133:1959–1967PubMedGoogle Scholar
  13. 13.
    Son M, Ghoreishi D, Ahn SJ, Burne RA, Hagen SJ (2015) Sharply tuned pH response of genetic competence regulation in Streptococcus mutans: a microfluidic study of the environmental sensitivity of comX. Appl Environ Microbiol 81:5622–5631CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Morrison DA, Khan R, Junges R, Amdal HA, Petersen FC (2015) Genome editing by natural genetic transformation in Streptococcus mutans. J Microbiol Methods 119:134–141CrossRefPubMedGoogle Scholar
  15. 15.
    Chang JC, LaSarre B, Jimenez JC, Aggarwal C, Federle MJ (2011) Two group A streptococcal peptide pheromones act through opposing Rgg regulators to control biofilm development. PLoS Pathog 7:e1002190CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Stevens KE, Chang D, Zwack EE, Sebert ME (2011) Competence in Streptococcus pneumoniae is regulated by the rate of ribosomal decoding errors. mBio 2:doi:  10.1128/mBio.00071-11
  17. 17.
    Lacks S, Hotchkiss RD (1960) A study of the genetic material determining an enzyme in Pneumococcus. Biochim Biophys Acta 39:508–518CrossRefPubMedGoogle Scholar
  18. 18.
    Reck M, Tomasch J, Wagner-Dobler I (2015) The alternative sigma factor SigX controls bacteriocin synthesis and competence, the two quorum sensing regulated traits in Streptococcus mutans. PLoS Genet 11, e1005353CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Gaustad P, Morrison DA (1998) Induction of transformation in streptococci by synthetic competence stimulating peptides. Methods Cell Sci 20:65–70CrossRefGoogle Scholar
  20. 20.
    Petersen FC, Pecharki D, Scheie AA (2004) Biofilm mode of growth of Streptococcus intermedius favored by a competence-stimulating signaling peptide. J Bacteriol 186:6327–6331CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Warren TK, Lund SA, Jones KF, Hruby DE (2007) Comparison of transformation protocols in Streptococcus gordonii and evaluation of native promoter strength using a multiple-copy plasmid. Can J Microbiol 53:417–426CrossRefPubMedGoogle Scholar
  22. 22.
    Palmer SR, Miller JH, Abranches J, Zeng L, Lefebure T, Richards VP, Lemos JA, Stanhope MJ, Burne RA (2013) Phenotypic heterogeneity of genomically-diverse isolates of Streptococcus mutans. PLoS One 8, e61358CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Ahn SJ, Wen ZT, Burne RA (2006) Multilevel control of competence development and stress tolerance in Streptococcus mutans UA159. Infect Immun 74:1631–1642CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Li YH, Lau PC, Lee JH, Ellen RP, Cvitkovitch DG (2001) Natural genetic transformation of Streptococcus mutans growing in biofilms. J Bacteriol 183:897–908CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Vickerman MM, Iobst S, Jesionowski AM, Gill SR (2007) Genome-wide transcriptional changes in Streptococcus gordonii in response to competence signaling peptide. J Bacteriol 189:7799–7807CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Kreth J, Merritt J, Shi W, Qi F (2005) Co-ordinated bacteriocin production and competence development: a possible mechanism for taking up DNA from neighbouring species. Mol Microbiol 57:392–404CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Petersen FC, Fimland G, Scheie AA (2006) Purification and functional studies of a potent modified quorum-sensing peptide and a two-peptide bacteriocin in Streptococcus mutans. Mol Microbiol 61:1322–1334CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Gabriela Salvadori
    • 1
  • Roger Junges
    • 1
  • Rabia Khan
    • 1
  • Heidi A. Åmdal
    • 1
  • Donald A. Morrison
    • 2
  • Fernanda C. Petersen
    • 1
    Email author
  1. 1.Department of Oral Biology, Faculty of DentistryUniversity of OsloBlindernNorway
  2. 2.Department of Biological Sciences, College of Liberal Arts and SciencesUniversity of Illinois at ChicagoChicagoUSA

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