Abstract
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.
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References
Federle MJ, Morrison DA (2012) One if by land, two if by sea: signalling to the ranks with CSP and XIP. Mol Microbiol 86:241–245
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 33:343–360
Johnsborg O, Eldholm V, Håvarstein LS (2007) Natural genetic transformation: prevalence, mechanisms and function. Res Microbiol 158:767–778
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 179:6589–6594
Morrison DA (1997) Streptococcal competence for genetic transformation: regulation by peptide pheromones. Microb Drug Resist 3:27–37
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 194:3781–3788
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 78589–78606
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 192:1444–1454
Monnet V, Juillard V, Gardan R (2016) Peptide conversations in Gram-positive bacteria. Crit Rev Microbiol 42:339–351
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 193:1863–1877
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
Chen JD, Morrison DA (1987) Modulation of competence for genetic transformation in Streptococcus pneumoniae. J Gen Microbiol 133:1959–1967
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 81:5622–5631
Morrison DA, Khan R, Junges R, Amdal HA, Petersen FC (2015) Genome editing by natural genetic transformation in Streptococcus mutans. J Microbiol Methods 119:134–141
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:e1002190
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
Lacks S, Hotchkiss RD (1960) A study of the genetic material determining an enzyme in Pneumococcus. Biochim Biophys Acta 39:508–518
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, e1005353
Gaustad P, Morrison DA (1998) Induction of transformation in streptococci by synthetic competence stimulating peptides. Methods Cell Sci 20:65–70
Petersen FC, Pecharki D, Scheie AA (2004) Biofilm mode of growth of Streptococcus intermedius favored by a competence-stimulating signaling peptide. J Bacteriol 186:6327–6331
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 53:417–426
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, e61358
Ahn SJ, Wen ZT, Burne RA (2006) Multilevel control of competence development and stress tolerance in Streptococcus mutans UA159. Infect Immun 74:1631–1642
Li YH, Lau PC, Lee JH, Ellen RP, Cvitkovitch DG (2001) Natural genetic transformation of Streptococcus mutans growing in biofilms. J Bacteriol 183:897–908
Vickerman MM, Iobst S, Jesionowski AM, Gill SR (2007) Genome-wide transcriptional changes in Streptococcus gordonii in response to competence signaling peptide. J Bacteriol 189:7799–7807
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 57:392–404
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 61:1322–1334
Acknowledgments
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).
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Salvadori, G., Junges, R., Khan, R., Ã…mdal, H.A., Morrison, D.A., Petersen, F.C. (2017). Natural Transformation of Oral Streptococci by Use of Synthetic Pheromones. In: Seymour, G., Cullinan, M., Heng, N. (eds) Oral Biology. Methods in Molecular Biology, vol 1537. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6685-1_13
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DOI: https://doi.org/10.1007/978-1-4939-6685-1_13
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