1887

Abstract

, a common species of the oral microbiome, expresses virulence genes promoting cariogenic dental biofilms, persistence in the bloodstream and cardiovascular infections.

Virulence gene expression is variable among strains and controlled by the transcription regulatory systems VicRK and CovR.

This study investigates polymorphisms in the and loci in strains isolated from the oral cavity or from the bloodstream, which were shown to differ in expression of , and downstream genes.

The transcriptional activities of , v and were compared by RT-qPCR between blood and oral strains after exposure to human serum. PCR-amplified promoter and/or coding regions of and of 18 strains (11 oral and 7 blood) were sequenced and compared to the reference strain UA159.

Serum exposure significantly reduced and / transcript levels in most strains (<0.05), but reductions were higher in oral than in blood strains. Single-nucleotide polymorphisms (SNPs) were detected in regulatory and coding regions, but SNPs affecting the CovR effector domain were only present in two blood strains. Although was highly conserved, showed several SNPs, and SNPs affecting VicK regions important for autokinase activity were found in three blood strains.

This study reveals transcriptional and structural diversity in and /, and identifies polymorphisms of functional relevance in blood strains, indicating that and might be important loci for adaptation to host selective pressures associated with virulence diversity.

Funding
This study was supported by the:
  • Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Award PNPD-2013)
    • Principle Award Recipient: ErikaN. Harth-Chu
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2015/22967-6)
    • Principle Award Recipient: LíviaA. Alves
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2017/19899-4)
    • Principle Award Recipient: LetíciaT. Oliveira
  • Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Award Code 001)
    • Principle Award Recipient: NotApplicable
  • Fundação de Amparo à Pesquisa do Estado de São Paulo (Award 2018/02054–4)
    • Principle Award Recipient: RenataO. Mattos-Graner
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001457
2021-12-23
2022-01-28
Loading full text...

Full text loading...

References

  1. Lemos JA, Palmer SR, Zeng L, Wen ZT, Kajfasz JK et al. The biology of Streptococcus mutans . Microbiol Spectr 2019; 7: [View Article]
    [Google Scholar]
  2. Mattos-Graner RO, Klein MI, Smith DJ. Lessons learned from clinical studies: roles of mutans streptococci in the pathogenesis of dental caries. Curr Oral Health Rep 2013; 1:70–78 [View Article]
    [Google Scholar]
  3. Nakano K, Nomura R, Matsumoto M, Ooshima T. Roles of oral bacteria in cardiovascular diseases — from molecular mechanisms to clinical cases: cell-surface structures of novel serotype k Streptococcus mutans strains and their correlation to virulence. J Pharmacol Sci 2010; 113:120–125 [View Article]
    [Google Scholar]
  4. Palmer SR, Miller JH, Abranches J, Zeng L, Lefebure T et al. Phenotypic heterogeneity of genomically-diverse isolates of Streptococcus mutans . PLoS One 2013; 8:e61358. [View Article] [PubMed]
    [Google Scholar]
  5. Avilés-Reyes A, Lemos JA, Abranches J. Collagen-binding proteins of Streptococcus mutans and related streptococci. Mol Oral Microbiol 2017; 32:89–106 [View Article] [PubMed]
    [Google Scholar]
  6. Hosoki S, Saito S, Tonomura S, Ishiyama H, Yoshimoto T et al. Oral carriage of Streptococcus mutans harboring the cnm gene relates to an increased incidence of cerebral microbleeds. Stroke 2020; 51:3632–3639 [View Article] [PubMed]
    [Google Scholar]
  7. Nomura R, Otsugu M, Naka S, Teramoto N, Kojima A et al. Contribution of the interaction of Streptococcus mutans serotype k strains with fibrinogen to the pathogenicity of infective endocarditis. Infect Immun 2014; 82:5223–5234 [View Article] [PubMed]
    [Google Scholar]
  8. Nomura R, Otsugu M, Hamada M, Matayoshi S, Teramoto N et al. Potential involvement of Streptococcus mutans possessing collagen binding protein Cnm in infective endocarditis. Sci Rep 2020; 10:19118. [View Article] [PubMed]
    [Google Scholar]
  9. Cornejo OE, Lefébure T, Bitar PDP, Lang P, Richards VP et al. Evolutionary and population genomics of the cavity causing bacteria Streptococcus mutans . Mol Biol Evol 2013; 30:881–893 [View Article] [PubMed]
    [Google Scholar]
  10. Argimón S, Caufield PW. Distribution of putative virulence genes in Streptococcus mutans strains does not correlate with caries experience. J Clin Microbiol 2011; 49:984–992 [View Article] [PubMed]
    [Google Scholar]
  11. Alves LA, Nomura R, Mariano FS, Harth-Chu EN, Stipp RN et al. CovR regulates Streptococcus mutans susceptibility to complement immunity and survival in blood. Infect Immun 2016; 84:3206–3219 [View Article] [PubMed]
    [Google Scholar]
  12. Alves LA, Harth-Chu EN, Palma TH, Stipp RN, Mariano FS et al. The two-component system VicRK regulates functions associated with Streptococcus mutans resistance to complement immunity. Mol Oral Microbiol 2017; 32:419–431 [View Article] [PubMed]
    [Google Scholar]
  13. Alves LA, Ganguly T, Harth-Chú ÉN, Kajfasz J, Lemos JA et al. PepO is a target of the two-component systems VicRK and CovR required for systemic virulence of Streptococcus mutans . Virulence 2020; 11:521–536 [View Article] [PubMed]
    [Google Scholar]
  14. Duque C, Stipp RN, Wang B, Smith DJ, Höfling JF et al. Downregulation of GbpB, a component of the VicRK regulon, affects biofilm formation and cell surface characteristics of Streptococcus mutans . Infect Immun 2011; 79:786–796 [View Article] [PubMed]
    [Google Scholar]
  15. Stipp RN, Gonçalves RB, Höfling JF, Smith DJ, Mattos-Graner RO. Transcriptional analysis of gtfB, gtfC, and gbpB and their putative response regulators in several isolates of Streptococcus mutans . Oral Microbiol Immunol 2008; 23:466–473 [View Article]
    [Google Scholar]
  16. Mattos-Graner RO, Jin S, King WF, Chen T, Smith DJ et al. Cloning of the Streptococcus mutans gene encoding glucan binding protein B and analysis of genetic diversity and protein production in clinical isolates. Infect Immun 2001; 69:6931–6941 [View Article] [PubMed]
    [Google Scholar]
  17. Senadheera MD, Guggenheim B, Spatafora GA, Huang Y-CC, 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 [View Article] [PubMed]
    [Google Scholar]
  18. Stipp RN, Boisvert H, Smith DJ, Höfling JF, Duncan MJ et al. CovR and VicRK regulate cell surface biogenesis genes required for biofilm formation in Streptococcus mutans . PLoS One 2013; 8:e58271. [View Article] [PubMed]
    [Google Scholar]
  19. Araújo Alves L, Ganguly T, Mattos-Graner RO, Kajfasz J, Harth-Chu EN et al. CovR and VicRKX regulate transcription of the collagen binding protein Cnm of Streptococcus mutans . J Bacteriol 2018; 200:e00141-18. [View Article] [PubMed]
    [Google Scholar]
  20. Ayala E, Downey JS, Mashburn-Warren L, Senadheera DB, Cvitkovitch DG et al. A biochemical characterization of the DNA binding activity of the response regulator VicR from Streptococcus mutans . PLoS One 2014; 9:e108027. [View Article] [PubMed]
    [Google Scholar]
  21. Bijlsma JJE, Groisman EA. Making informed decisions: regulatory interactions between two-component systems. Trends in Microbiology 2003; 11:359–366 [View Article]
    [Google Scholar]
  22. Mattos-Graner RO, Duncan MJ. Two-component signal transduction systems in oral bacteria. J Oral Microbiol 2017; 9:1400858. [View Article] [PubMed]
    [Google Scholar]
  23. Senadheera DB, Cordova M, Ayala EA, Chávez de Paz LE, Singh K et al. Regulation of bacteriocin production and cell death by the VicRK signaling system in Streptococcus mutans . J Bacteriol 2012; 194:1307–1316 [View Article] [PubMed]
    [Google Scholar]
  24. Downey JS, Mashburn-Warren L, Ayala EA, Senadheera DB, Hendrickson WK et al. In vitro manganese-dependent cross-talk between Streptococcus mutans VicK and GcrR: implications for overlapping stress response pathways. PLoS One 2014; 9:e115975. [View Article] [PubMed]
    [Google Scholar]
  25. Gryllos I, Tran-Winkler HJ, Cheng M-F, Chung H, Bolcome R 3rd et al. Induction of group A Streptococcus virulence by a human antimicrobial peptide. Proc Natl Acad Sci U S A 2008; 105:16755–16760 [View Article] [PubMed]
    [Google Scholar]
  26. Khara P, Mohapatra SS, Biswas I. Role of CovR phosphorylation in gene transcription in Streptococcus mutans . Microbiology (Reading) 2018; 164:704–715 [View Article] [PubMed]
    [Google Scholar]
  27. Biswas S, Biswas I. Regulation of the glucosyltransferase (gtfBC) operon by CovR in Streptococcus mutans . J Bacteriol 2006; 188:988–998 [View Article] [PubMed]
    [Google Scholar]
  28. Mattos-Graner RO, Smith DJ, King WF, Mayer MPA. Water-insoluble glucan synthesis by mutans Streptococcal strains correlates with caries incidence in 12- to 30-month-old children. J Dent Res 2016; 79:1371–1377 [View Article]
    [Google Scholar]
  29. Nakano K, Lapirattanakul J, Nomura R, Nemoto H, Alaluusua S et al. Streptococcus mutans clonal variation revealed by multilocus sequence typing. J Clin Microbiol 2007; 45:2616–2625 [View Article] [PubMed]
    [Google Scholar]
  30. Ajdić D, McShan WM, McLaughlin RE, Savić G, Chang J et al. Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc Natl Acad Sci U S A 2002; 99:14434–14439 [View Article] [PubMed]
    [Google Scholar]
  31. Madeira F, Park YM, Lee J, Buso N, Gur T et al. The EMBL-EBI search and sequence analysis tools APIs in 2019. Nucleic Acids Res 2019; 47:W636–W641 [View Article] [PubMed]
    [Google Scholar]
  32. Letunic I, Bork P. 20 years of the SMART protein domain annotation resource. Nucleic Acids Res 2018; 46:D493–D496 [View Article] [PubMed]
    [Google Scholar]
  33. Letunic I, Khedkar S, Bork P. SMART: recent updates, new developments and status in 2020. Nucleic Acids Res 2021; 49:D458–D460 [View Article] [PubMed]
    [Google Scholar]
  34. Lemoine F, Correia D, Lefort V, Doppelt-Azeroual O, Mareuil F et al. NGPhylogeny.fr: new generation phylogenetic services for non-specialists. Nucleic Acids Res 2019; 47:W260–W265 [View Article] [PubMed]
    [Google Scholar]
  35. Chong P, Drake L, Biswas I. Modulation of covR expression in Streptococcus mutans UA159. J Bacteriol 2008; 190:4478–4488 [View Article] [PubMed]
    [Google Scholar]
  36. Adler CJ, Dobney K, Weyrich LS, Kaidonis J, Walker AW et al. Sequencing ancient calcified dental plaque shows changes in oral microbiota with dietary shifts of the Neolithic and Industrial revolutions. Nat Genet 2013; 45:450–455 [View Article] [PubMed]
    [Google Scholar]
  37. Achtman M, Zhou Z. Metagenomics of the modern and historical human oral microbiome with phylogenetic studies on Streptococcus mutans and Streptococcus sobrinus . Philos Trans R Soc Lond B Biol Sci 2020; 375:20190573. [View Article] [PubMed]
    [Google Scholar]
  38. Avilés-Reyes A, Miller JH, Simpson-Haidaris PJ, Lemos JA, Abranches J. Cnm is a major virulence factor of invasive Streptococcus mutans and part of a conserved three-gene locus. Mol oral Microbiol 2014; 29:11–23 [View Article]
    [Google Scholar]
  39. Napimoga MH, Kamiya RU, Rosa RT, Rosa EAntonioR, Höfling JF et al. Genotypic diversity and virulence traits of Streptococcus mutans in caries-free and caries-active individuals. J Med Microbiol 2004; 53:697–703 [View Article]
    [Google Scholar]
  40. Mattos-Graner RO, Napimoga MH, Fukushima K, Duncan MJ, Smith DJ. Comparative analysis of Gtf isozyme production and diversity in isolates of Streptococcus mutans with different biofilm growth phenotypes. J Clin Microbiol 2004; 42:4586–4592 [View Article] [PubMed]
    [Google Scholar]
  41. Bedoya-Correa CM, Rincón Rodríguez RJ, Parada-Sanchez MT. Genomic and phenotypic diversity of Streptococcus mutans . J Oral Biosci 2019; 61:22–31 [View Article]
    [Google Scholar]
  42. Engleberg NC, Heath A, Miller A, Rivera C, DiRita VJ. Spontaneous mutations in the CsrRS two‐component regulatory system of Streptococcus pyogenes result in enhanced virulence in a murine model of skin and soft tissue infection. J Infect Dis 2001; 183:1043–1054 [View Article]
    [Google Scholar]
  43. Walker MJ, Hollands A, Sanderson-Smith ML, Cole JN, Kirk JK et al. DNase Sda1 provides selection pressure for a switch to invasive group A streptococcal infection. Nat Med 2007; 13:981–985 [View Article]
    [Google Scholar]
  44. Ato M, Ikebe T, Kawabata H, Takemori T, Watanabe H. Incompetence of neutrophils to invasive group A streptococcus is attributed to induction of plural virulence factors by dysfunction of a regulator. PLoS One 2008; 3:e3455. [View Article] [PubMed]
    [Google Scholar]
  45. Sumby P, Whitney AR, Graviss EA, DeLeo FR, Musser JM. Genome-wide analysis of group a streptococci reveals a mutation that modulates global phenotype and disease specificity. PLoS Pathog 2006; 2:e5. [View Article] [PubMed]
    [Google Scholar]
  46. Horstmann N, Sahasrabhojane P, Suber B, Kumaraswami M, Olsen RJ et al. Distinct single amino acid replacements in the control of virulence regulator protein differentially impact streptococcal pathogenesis. PLoS Pathog 2011; 7:e1002311. [View Article] [PubMed]
    [Google Scholar]
  47. Friães A, Pato C, Melo-Cristino J, Ramirez M. Consequences of the variability of the CovRS and RopB regulators among Streptococcus pyogenes causing human infections. Sci Rep 2015; 5:12057. [View Article] [PubMed]
    [Google Scholar]
  48. Otsuji K, Fukuda K, Maruoka T, Ogawa M, Saito M. Acquisition of genetic mutations in Group A Streptococci at infection site and subsequent systemic dissemination of the mutants with lethal mutations in a streptococcal toxic shock syndrome mouse model. Microbial Pathogenesis 2020; 143:104116 [View Article]
    [Google Scholar]
  49. Wilkening RV, Federle MJ. Evolutionary constraints shaping Streptococcus pyogenes-host interactions. Trends Microbiol 2017; 25:562–572 [View Article] [PubMed]
    [Google Scholar]
  50. Miyoshi‐Akiyama T, Ikebe T, Watanabe H, Uchiyama T, Kirikae T et al. Use of DNA arrays to identify a mutation in the negative regulator, csrR, responsible for the high virulence of a naturally occurring type M3 group A Streptococcus clinical isolate. J INFECT DIS 2006; 193:1677–1684 [View Article]
    [Google Scholar]
  51. Ikebe T, Ato M, Matsumura T, Hasegawa H, Sata T et al. Highly frequent mutations in negative regulators of multiple virulence genes in group A streptococcal toxic shock syndrome isolates. PLoS Pathog 2010; 6:e1000832. [View Article] [PubMed]
    [Google Scholar]
  52. Churchward G. The two faces of Janus: virulence gene regulation by CovR/S in group A streptococci. Mol Microbiol 2007; 64:34–41 [View Article]
    [Google Scholar]
  53. Gao J, Gusa AA, Scott JR, Churchward G. Binding of the global response regulator protein CovR to the sag promoter of Streptococcus pyogenes reveals a new mode of CovR-DNA Interaction. Journal of Biological Chemistry 2005; 280:38948–38956 [View Article]
    [Google Scholar]
  54. Buckley SJ, Timms P, Davies MR, McMillan DJ. In silico characterisation of the two-component system regulators of Streptococcus pyogenes . PLoS One 2018; 13:e0199163. [View Article] [PubMed]
    [Google Scholar]
  55. Dubrac S, Bisicchia P, Devine KM, Msadek T. A matter of life and death: cell wall homeostasis and the WalKR (YycGF) essential signal transduction pathway. Mol Microbiol 2008; 70:1307–1322 [View Article]
    [Google Scholar]
  56. Wayne KJ, Li S, Kazmierczak KM, Tsui HCT, Winkler ME. Involvement of WalK (VicK) phosphatase activity in setting WalR (VicR) response regulator phosphorylation level and limiting cross-talk in Streptococcus pneumoniae D39 cells. Mol Microbiol 2012; 86:645–660 [View Article] [PubMed]
    [Google Scholar]
  57. Sham LT, Barendt SM, Kopecky KE, Winkler ME. Essential PcsB putative peptidoglycan hydrolase interacts with the essential FtsXSpn cell division protein in Streptococcus pneumoniae D39. Proc Natl Acad Sci U S A 2011; 108:E1061–9 [View Article] [PubMed]
    [Google Scholar]
  58. Wang Y, Cao W, Merritt J, Xie Z, Liu H. Characterization of FtsH essentiality in Streptococcus mutans via genetic suppression. Front Genet 2021; 12:659220. [View Article] [PubMed]
    [Google Scholar]
  59. Zhuang PL, Yu LX, Liao JK, Zhou Y, Lin HC. Relationship between the genetic polymorphisms of vicR and vicK Streptococcus mutans genes and early childhood caries in two-year-old children. BMC Oral Health 2018; 18:39. [View Article] [PubMed]
    [Google Scholar]
  60. Senadheera MD, Lee AWC, Hung DCI, Spatafora GA, Goodman SD et al. The Streptococcus mutans vicX gene product modulates gtfB/C expression, biofilm formation, genetic competence, and oxidative stress tolerance. J Bacteriol 2007; 189:1451–1458 [View Article] [PubMed]
    [Google Scholar]
  61. Lei L, Stipp RN, Chen T, Wu SZ, Hu T et al. Activity of Streptococcus mutans VicR is modulated by antisense RNA. J Dent Res 2018; 97:1477–1484 [View Article] [PubMed]
    [Google Scholar]
  62. Wang S, Long L, Yang X, Qiu Y, Tao T et al. Dissecting the role of vick phosphatase in aggregation and biofilm formation of Streptococcus mutans . J Dent Res 2021; 100:631–638 [View Article]
    [Google Scholar]
  63. Wang C, Sang J, Wang J, Su M, Downey JS et al. Mechanistic insights revealed by the crystal structure of a histidine kinase with signal transducer and sensor domains. PLoS Biol 2013; 11:e1001493. [View Article] [PubMed]
    [Google Scholar]
  64. Lu J, Cheng L, Huang Y, Jiang Y, Chu C-H et al. Resumptive Streptococcus mutans persisters induced from dimethylaminododecyl methacrylate elevated the cariogenic virulence by up-regulating the quorum-sensing and VicRK pathway genes. Front Microbiol 2019; 10:3102. [View Article] [PubMed]
    [Google Scholar]
  65. Filho JG, Vizoto NL, Dias de Sena M, Mendes da Camara D. Genetic and physiological effects of subinhibitory concentrations of oral antimicrobial agents on Streptococcus mutans biofilms: Effects of sodium fluoride and chlorhexidine on Streptococcus mutans . Microb Pathog 2021; 150: [View Article]
    [Google Scholar]
  66. Friedman L, Alder JD, Silverman JA. Genetic changes that correlate with reduced susceptibility to daptomycin in Staphylococcus aureus . Antimicrob Agents Chemother 2006; 50:2137–2145 [View Article] [PubMed]
    [Google Scholar]
  67. Howden BP, McEvoy CRE, Allen DL, Chua K, Gao W et al. Evolution of multidrug resistance during Staphylococcus aureus infection involves mutation of the essential two component regulator WalKR. PLoS Pathog 2011; 7:e1002359. [View Article] [PubMed]
    [Google Scholar]
  68. Werth BJ, Ashford NK, Penewit K, Waalkes A, Holmes EA et al. Dalbavancin exposure in vitro selects for dalbavancin-non-susceptible and vancomycin-intermediate strains of methicillin-resistant Staphylococcus aureus . Clin Microbiol Infect 2021; 27:910 [View Article] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.001457
Loading
/content/journal/jmm/10.1099/jmm.0.001457
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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