1887

Abstract

Purpose. The objective of the present study was to investigate whether Streptococcus sanguinis SpxA2 plays a role in competence development and endogenous H2O2 generation, and whether the SpxA2 Cys10-XX-Cys13 (CXXC) motif is involved in competence development.

Methodology. The competence development of wild-type S. sanguinis (SK36) and its derivatives was compared by transformation efficiency assay and real-time RT-PCR. The spx allele mutants, spxA2 (C10A) and spxA2 (C13A), were constructed by site-directed mutagenesis. The Δpox mutant was treated with 1 mM H2O2 to exclude the effect of other Pox products on competence development.

Results. Compared with the wild-type (4.42±0.58×10), the ΔspxA2 mutant showed decreased transformation efficiency (0.07±0.03×10). Furthermore, there was a 2- to 15-fold reduction in ΔspxA2 mutant com gene expression. SpxA2 was able to down-regulate endogenous H2O2 generation by repressing pox expression. Additionally, endogenous H2O2 negatively regulated competence without affecting spxA2 expression. The Δpox mutant increased com gene expression (2- to 8-fold), but the 1 mM H2O2-treated Δpox mutant showed decreased com gene expression. Interestingly, the ΔspxA2Δpox mutant showed enhanced competence-associated parameters. The fact that spxA2 (C10A) and spxA2 (C13A) behaved like the ΔspxA2 mutant revealed the role of the CXXC motif in competence development.

Conclusion. Although the intricate relationship between SpxA2, pox-mediated H2O2 production and competence development was clarified in S. sanguinis, it would be worthwhile to explore further whether H2O2 is involved in competence development through oxidizing the SpxA2 CXXC motif.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000506
2017-07-10
2019-10-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/66/7/981.html?itemId=/content/journal/jmm/10.1099/jmm.0.000506&mimeType=html&fmt=ahah

References

  1. Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE. Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 2005;43:5721–5732 [CrossRef][PubMed]
    [Google Scholar]
  2. Socransky SS, Manganiello AD, Propas D, Oram V, van Houte J. Bacteriological studies of developing supragingival dental plaque. J Periodontal Res 1977;12:90–106 [CrossRef][PubMed]
    [Google Scholar]
  3. Stingu CS, Eschrich K, Rodloff AC, Schaumann R, Jentsch H. Periodontitis is associated with a loss of colonization by Streptococcus sanguinis. J Med Microbiol 2008;57:495–499 [CrossRef][PubMed]
    [Google Scholar]
  4. Di Filippo S, Delahaye F, Semiond B, Celard M, Henaine R et al. Current patterns of infective endocarditis in congenital heart disease. Heart 2006;92:1490–1495 [CrossRef][PubMed]
    [Google Scholar]
  5. Lemos JA, Abranches J, Burne RA. Responses of cariogenic streptococci to environmental stresses. Curr Issues Mol Biol 2005;7:95–107[PubMed]
    [Google Scholar]
  6. Monds RD, O'Toole GA. The developmental model of microbial biofilms: ten years of a paradigm up for review. Trends Microbiol 2009;17:73–87 [CrossRef][PubMed]
    [Google Scholar]
  7. Sherry L, Ramage G, Kean R, Borman A, Johnson EM et al. Biofilm-forming capability of highly virulent, multidrug-resistant Candida auris. Emerg Infect Dis 2017;23:328–331 [CrossRef][PubMed]
    [Google Scholar]
  8. 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]
  9. Li YH, Tang N, Aspiras MB, Lau PC, Lee JH et al. A quorum-sensing signaling system essential for genetic competence in Streptococcus mutans is involved in biofilm formation. J Bacteriol 2002;184:2699–2708[PubMed][CrossRef]
    [Google Scholar]
  10. Loo CY, Corliss DA, Ganeshkumar N. Streptococcus gordonii biofilm formation: identification of genes that code for biofilm phenotypes. J Bacteriol 2000;182:1374–1382 [CrossRef][PubMed]
    [Google Scholar]
  11. Claverys JP, Prudhomme M, Martin B. Induction of competence regulons as a general response to stress in gram-positive bacteria. Annu Rev Microbiol 2006;60:451–475 [CrossRef][PubMed]
    [Google Scholar]
  12. Dufour D, Lévesque CM. Bacterial behaviors associated with the quorum-sensing peptide pheromone ('alarmone') in streptococci. Future Microbiol 2013;8:593–605 [CrossRef][PubMed]
    [Google Scholar]
  13. Suntharalingam P, Cvitkovitch DG. Quorum sensing in streptococcal biofilm formation. Trends Microbiol 2005;13:3–6 [CrossRef][PubMed]
    [Google Scholar]
  14. Ribbe J, Maier B. Density-dependent differentiation of bacteria in spatially structured open systems. Biophys J 2016;110:1648–1660 [CrossRef][PubMed]
    [Google Scholar]
  15. Charpentier X, Polard P, Claverys JP. Induction of competence for genetic transformation by antibiotics: convergent evolution of stress responses in distant bacterial species lacking SOS?. Curr Opin Microbiol 2012;15:570–576 [CrossRef][PubMed]
    [Google Scholar]
  16. Didelot X, Maiden MC. Impact of recombination on bacterial evolution. Trends Microbiol 2010;18:315–322 [CrossRef][PubMed]
    [Google Scholar]
  17. 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]
  18. Nakano S, Küster-Schöck E, Grossman AD, Zuber P. Spx-dependent global transcriptional control is induced by thiol-specific oxidative stress in Bacillus subtilis. Proc Natl Acad Sci USA 2003;100:13603–13608 [CrossRef][PubMed]
    [Google Scholar]
  19. Newberry KJ, Nakano S, Zuber P, Brennan RG. Crystal structure of the Bacillus subtilis anti-alpha, global transcriptional regulator, Spx, in complex with the α C-terminal domain of RNA polymerase. Proc Natl Acad Sci USA 2005;102:15839–15844 [CrossRef][PubMed]
    [Google Scholar]
  20. Zuber P. Spx-RNA polymerase interaction and global transcriptional control during oxidative stress. J Bacteriol 2004;186:1911–1918 [CrossRef][PubMed]
    [Google Scholar]
  21. Nakano S, Erwin KN, Ralle M, Zuber P. Redox-sensitive transcriptional control by a thiol/disulphide switch in the global regulator, Spx. Mol Microbiol 2005;55:498–510 [CrossRef][PubMed]
    [Google Scholar]
  22. Nakano MM, Nakano S, Zuber P. Spx (YjbD), a negative effector of competence in Bacillus subtilis, enhances ClpC–MecA–ComK interaction. Mol Microbiol 2002;44:1341–1349 [CrossRef][PubMed]
    [Google Scholar]
  23. Turlan C, Prudhomme M, Fichant G, Martin B, Gutierrez C. SpxA1, a novel transcriptional regulator involved in X-state (competence) development in Streptococcus pneumoniae. Mol Microbiol 2009;73:492–506 [CrossRef][PubMed]
    [Google Scholar]
  24. Chen L, Ge X, Wang X, Patel JR, Xu P. SpxA1 involved in hydrogen peroxide production, stress tolerance and endocarditis virulence in Streptococcus sanguinis. PLoS One 2012;7:e40034 [CrossRef][PubMed]
    [Google Scholar]
  25. Galvão LC, Miller JH, Kajfasz JK, Scott-Anne K, Freires IA et al. Transcriptional and phenotypic characterization of novel Spx-regulated genes in Streptococcus mutans. PLoS One 2015;10:e0124969 [CrossRef][PubMed]
    [Google Scholar]
  26. Bättig P, Mühlemann K. Influence of the spxB gene on competence in Streptococcus pneumoniae. J Bacteriol 2008;190:1184–1189 [CrossRef][PubMed]
    [Google Scholar]
  27. Carlsson J, Edlund MB, Lundmark SK. Characteristics of a hydrogen peroxide-forming pyruvate oxidase from Streptococcus sanguis. Oral Microbiol Immunol 1987;2:15–20 [CrossRef][PubMed]
    [Google Scholar]
  28. Brynildsen MP, Winkler JA, Spina CS, Macdonald IC, Collins JJ. Potentiating antibacterial activity by predictably enhancing endogenous microbial ROS production. Nat Biotechnol 2013;31:160–165 [CrossRef][PubMed]
    [Google Scholar]
  29. Dwyer DJ, Camacho DM, Kohanski MA, Callura JM, Collins JJ. Antibiotic-induced bacterial cell death exhibits physiological and biochemical hallmarks of apoptosis. Mol Cell 2012;46:561–572 [CrossRef][PubMed]
    [Google Scholar]
  30. Foti JJ, Devadoss B, Winkler JA, Collins JJ, Walker GC. Oxidation of the guanine nucleotide pool underlies cell death by bactericidal antibiotics. Science 2012;336:315–319 [CrossRef][PubMed]
    [Google Scholar]
  31. Charpentier X, Kay E, Schneider D, Shuman HA. Antibiotics and UV radiation induce competence for natural transformation in Legionella pneumophila. J Bacteriol 2011;193:1114–1121 [CrossRef][PubMed]
    [Google Scholar]
  32. Saito M, Seki M, Iida K, Nakayama H, Yoshida S. A novel agar medium to detect hydrogen peroxide-producing bacteria based on the prussian blue-forming reaction. Microbiol Immunol 2007;51:889–892 [CrossRef][PubMed]
    [Google Scholar]
  33. Li T, Zhai S, Xu M, Shang M, Gao Y et al. SpxB-mediated H2O2 induces programmed cell death in Streptococcus sanguinis. J Basic Microbiol 2016;56:741–752 [CrossRef][PubMed]
    [Google Scholar]
  34. Zheng L, Chen Z, Itzek A, Ashby M, Kreth J. Catabolite control protein A controls hydrogen peroxide production and cell death in Streptococcus sanguinis. J Bacteriol 2011;193:516–526 [CrossRef][PubMed]
    [Google Scholar]
  35. Xu P, Alves JM, Kitten T, Brown A, Chen Z et al. Genome of the opportunistic pathogen Streptococcus sanguinis. J Bacteriol 2007;189:3166–3175 [CrossRef][PubMed]
    [Google Scholar]
  36. Pestova EV, Håvarstein LS, Morrison DA. Regulation of competence for genetic transformation in Streptococcus pneumoniae by an auto-induced peptide pheromone and a two-component regulatory system. Mol Microbiol 1996;21:853–862 [CrossRef][PubMed]
    [Google Scholar]
  37. Rodriguez AM, Callahan JE, Fawcett P, Ge X, Xu P et al. Physiological and molecular characterization of genetic competence in Streptococcus sanguinis. Mol Oral Microbiol 2011;26:99–116 [CrossRef][PubMed]
    [Google Scholar]
  38. Luo P, Li H, Morrison DA. ComX is a unique link between multiple quorum sensing outputs and competence in Streptococcus pneumoniae. Mol Microbiol 2003;50:623–633 [CrossRef][PubMed]
    [Google Scholar]
  39. Luo P, Morrison DA. Transient association of an alternative sigma factor, ComX, with RNA polymerase during the period of competence for genetic transformation in Streptococcus pneumoniae. J Bacteriol 2003;185:349–358 [CrossRef][PubMed]
    [Google Scholar]
  40. Liu J, Zuber P. The ClpX protein of Bacillus subtilis indirectly influences RNA polymerase holoenzyme composition and directly stimulates σH-dependent transcription. Mol Microbiol 2000;37:885–897 [CrossRef][PubMed]
    [Google Scholar]
  41. Laurenceau R, Péhau-Arnaudet G, Baconnais S, Gault J, Malosse C et al. A type IV pilus mediates DNA binding during natural transformation in Streptococcus pneumoniae. PLoS Pathog 2013;9:e1003473 [CrossRef][PubMed]
    [Google Scholar]
  42. Metzger LC, Blokesch M. Composition of the DNA-uptake complex of Vibrio cholerae. Mob Genet Elements 2014;4:e28142 [CrossRef][PubMed]
    [Google Scholar]
  43. Fontaine L, Wahl A, Fléchard M, Mignolet J, Hols P. Regulation of competence for natural transformation in streptococci. Infect Genet Evol 2015;33:343–360 [CrossRef][PubMed]
    [Google Scholar]
  44. Pericone CD, Bae D, Shchepetov M, Mccool T, Weiser JN. Short-sequence tandem and nontandem DNA repeats and endogenous hydrogen peroxide production contribute to genetic instability of Streptococcus pneumoniae. J Bacteriol 2002;184:4392–4399 [CrossRef][PubMed]
    [Google Scholar]
  45. Pericone CD, Park S, Imlay JA, Weiser JN. Factors contributing to hydrogen peroxide resistance in Streptococcus pneumoniae include pyruvate oxidase (SpxB) and avoidance of the toxic effects of the fenton reaction. J Bacteriol 2003;185:6815–6825 [CrossRef][PubMed]
    [Google Scholar]
  46. 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/jmm/10.1099/jmm.0.000506
Loading
/content/journal/jmm/10.1099/jmm.0.000506
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