1887

Abstract

The Streptococcus mutans Cid/Lrg system represents an ideal model for studying this organism’s ability to withstand various stressors encountered in the oral cavity. The lrg and cid operons display distinct and opposite patterns of expression in response to growth phase and glucose levels, suggesting that the activity and regulation of these proteins must be tightly coordinated in the cell and closely associated with metabolic pathways of the organism. Here, we demonstrate that expression of the cid and lrg operons is directly mediated by a global transcriptional regulator CcpA in response to glucose levels. Comparison of the cid and lrg promoter regions with the conserved CcpA binding motif revealed the presence of two potential cre sites (for CcpA binding) in the cid promoter (designated cid-cre1 and cid-cre2), which were arranged in a similar manner to those previously identified in the lrg promoter region (designated lrg-cre1 and lrg-cre2). We demonstrated that CcpA binds to both the cid and lrg promoters with a high affinity, but has an opposing glucose-dependent effect on the regulation of cid (positive) and lrg (negative) expression. DNase I footprinting analyses revealed potential binding sequences for CcpA in both cid and lrg promoter regions. Collectively, these data suggest that CcpA is a direct regulator of cid and lrg expression, and are suggestive of a potential mechanism by which Cid/Lrg-mediated virulence and cellular homeostasis is integrated with signals associated with both the environment and cellular metabolic status.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000744
2018-11-26
2019-10-23
Loading full text...

Full text loading...

References

  1. Loesche WJ. Role of Streptococcus mutans in human dental decay. Microbiol Rev 1986;50:353–380[PubMed]
    [Google Scholar]
  2. Burne RA, Ahn SJ, Wen ZT, Zeng L, Lemos JA et al. Opportunities for disrupting cariogenic biofilms. Adv Dent Res 2009;21:17–20 [CrossRef][PubMed]
    [Google Scholar]
  3. Burne RA, Zeng L, Ahn SJ, Palmer SR, Liu Y et al. Progress dissecting the oral microbiome in caries and health. Adv Dent Res 2012;24:77–80 [CrossRef][PubMed]
    [Google Scholar]
  4. Bowen WH, Burne RA, Wu H, Koo H. Oral biofilms: pathogens, matrix, and polymicrobial interactions in microenvironments. Trends Microbiol 2018;26: [CrossRef][PubMed]
    [Google Scholar]
  5. Bayles KW. Are the molecular strategies that control apoptosis conserved in bacteria?. Trends Microbiol 2003;11:306–311 [CrossRef][PubMed]
    [Google Scholar]
  6. Bayles KW. The biological role of death and lysis in biofilm development. Nat Rev Microbiol 2007;5:721–726 [CrossRef][PubMed]
    [Google Scholar]
  7. Bayles KW. Bacterial programmed cell death: making sense of a paradox. Nat Rev Microbiol 2014;12:63–69 [CrossRef][PubMed]
    [Google Scholar]
  8. Ahn SJ, Gu T, Koh J, Rice KC. Remodeling of the Streptococcus mutans proteome in response to LrgAB and external stresses. Sci Rep 2017;7:14063 [CrossRef][PubMed]
    [Google Scholar]
  9. Ahn SJ, Rice KC, Oleas J, Bayles KW, Burne RA. The Streptococcus mutans Cid and Lrg systems modulate virulence traits in response to multiple environmental signals. Microbiology 2010;156:3136–3147 [CrossRef][PubMed]
    [Google Scholar]
  10. Rice KC, Turner ME, Carney OV, Gu T, Ahn SJ. Modification of the Streptococcus mutans transcriptome by LrgAB and environmental stressors. Microb Genom 2017;3:e000104 [CrossRef][PubMed]
    [Google Scholar]
  11. Ahn SJ, Qu MD, Roberts E, Burne RA, Rice KC. Identification of the Streptococcus mutans LytST two-component regulon reveals its contribution to oxidative stress tolerance. BMC Microbiol 2012;12:187 [CrossRef][PubMed]
    [Google Scholar]
  12. Ahn SJ, Rice KC. Understanding the Streptococcus mutans Cid/Lrg System through CidB Function. Appl Environ Microbiol 2016;82:6189–6203 [CrossRef][PubMed]
    [Google Scholar]
  13. Young R. Bacteriophage lysis: mechanism and regulation. Microbiol Rev 1992;56:430–481[PubMed]
    [Google Scholar]
  14. Young R. Bacteriophage holins: deadly diversity. J Mol Microbiol Biotechnol 2002;4:21–36[PubMed]
    [Google Scholar]
  15. Young R, Bläsi U. Holins: form and function in bacteriophage lysis. FEMS Microbiol Rev 1995;17:191–205 [CrossRef][PubMed]
    [Google Scholar]
  16. Bayles KW. The bactericidal action of penicillin: new clues to an unsolved mystery. Trends Microbiol 2000;8:274–278 [CrossRef][PubMed]
    [Google Scholar]
  17. Rice KC, Bayles KW. Molecular control of bacterial death and lysis. Microbiol Mol Biol Rev 2008;72:85–109 [CrossRef][PubMed]
    [Google Scholar]
  18. Shemesh M, Tam A, Kott-Gutkowski M, Feldman M, Steinberg D. DNA-microarrays identification of Streptococcus mutans genes associated with biofilm thickness. BMC Microbiol 2008;8:236 [CrossRef][PubMed]
    [Google Scholar]
  19. Groicher KH, Firek BA, Fujimoto DF, Bayles KW. The Staphylococcus aureus lrgAB operon modulates murein hydrolase activity and penicillin tolerance. J Bacteriol 2000;182:1794–1801 [CrossRef][PubMed]
    [Google Scholar]
  20. Rice KC, Bayles KW. Death's toolbox: examining the molecular components of bacterial programmed cell death. Mol Microbiol 2003;50:729–738 [CrossRef][PubMed]
    [Google Scholar]
  21. Rice KC, Firek BA, Nelson JB, Yang SJ, Patton TG et al. The Staphylococcus aureus cidAB operon: evaluation of its role in regulation of murein hydrolase activity and penicillin tolerance. J Bacteriol 2003;185:2635–2643 [CrossRef][PubMed]
    [Google Scholar]
  22. Henkin TM. The role of CcpA transcriptional regulator in carbon metabolism in Bacillus subtilis. FEMS Microbiol Lett 1996;135:9–15 [CrossRef][PubMed]
    [Google Scholar]
  23. Deutscher J. The mechanisms of carbon catabolite repression in bacteria. Curr Opin Microbiol 2008;11:87–93 [CrossRef][PubMed]
    [Google Scholar]
  24. Görke B, Stülke J. Carbon catabolite repression in bacteria: many ways to make the most out of nutrients. Nat Rev Microbiol 2008;6:613–624 [CrossRef][PubMed]
    [Google Scholar]
  25. Miwa Y, Nakata A, Ogiwara A, Yamamoto M, Fujita Y. Evaluation and characterization of catabolite-responsive elements (cre) of Bacillus subtilis. Nucleic Acids Res 2000;28:1206–1210 [CrossRef][PubMed]
    [Google Scholar]
  26. Abranches J, Nascimento MM, Zeng L, Browngardt CM, Wen ZT et al. CcpA regulates central metabolism and virulence gene expression in Streptococcus mutans. J Bacteriol 2008;190:2340–2349 [CrossRef][PubMed]
    [Google Scholar]
  27. Sonenshein AL. Control of key metabolic intersections in Bacillus subtilis. Nat Rev Microbiol 2007;5:917–927 [CrossRef][PubMed]
    [Google Scholar]
  28. Deutscher J, Küster E, Bergstedt U, Charrier V, Hillen W. Protein kinase-dependent HPr/CcpA interaction links glycolytic activity to carbon catabolite repression in Gram-positive bacteria. Mol Microbiol 1995;15:1049–1053 [CrossRef][PubMed]
    [Google Scholar]
  29. Deutscher J, Saier MH. ATP-dependent protein kinase-catalyzed phosphorylation of a seryl residue in HPr, a phosphate carrier protein of the phosphotransferase system in Streptococcus pyogenes. Proc Natl Acad Sci USA 1983;80:6790–6794 [CrossRef][PubMed]
    [Google Scholar]
  30. Saier MH. Cyclic AMP-independent catabolite repression in bacteria. FEMS Microbiol Lett 1996;138:97–103 [CrossRef][PubMed]
    [Google Scholar]
  31. Terleckyj B, Shockman GD. Amino acid requirements of Streptococcus mutans and other oral streptococci. Infect Immun 1975;11:656–664[PubMed]
    [Google Scholar]
  32. Son M, Ahn SJ, Guo Q, Burne RA, Hagen SJ. Microfluidic study of competence regulation in Streptococcus mutans: environmental inputs modulate bimodal and unimodal expression of comX. Mol Microbiol 2012;86:258–272 [CrossRef][PubMed]
    [Google Scholar]
  33. Son M, Ghoreishi D, Ahn SJ, Burne RA, Hagen SJ. Sharply tuned pH Response of genetic competence regulation in Streptococcus mutans: a microfluidic study of the environmental sensitivity of comX. Appl Environ Microbiol 2015;81:5622–5631 [CrossRef][PubMed]
    [Google Scholar]
  34. Wen ZT, Burne RA. Analysis of cis- and trans-acting factors involved in regulation of the Streptococcus mutans fructanase gene (fruA). J Bacteriol 2002;184:126–133 [CrossRef][PubMed]
    [Google Scholar]
  35. Kim JN, Burne RA. CcpA and CodY coordinate acetate metabolism in Streptococcus mutans. Appl Environ Microbiol 2017;83: [CrossRef][PubMed]
    [Google Scholar]
  36. Zeng L, Dong Y, Burne RA. Characterization of cis-acting sites controlling arginine deiminase gene expression in Streptococcus gordonii. J Bacteriol 2006;188:941–949 [CrossRef][PubMed]
    [Google Scholar]
  37. Yindeeyoungyeon W, Schell MA. Footprinting with an automated capillary DNA sequencer. Biotechniques 2000;29:1034–1036, 1038, 1040–1031 [CrossRef][PubMed]
    [Google Scholar]
  38. de Been M, Bart MJ, Abee T, Siezen RJ, Francke C. The identification of response regulator-specific binding sites reveals new roles of two-component systems in Bacillus cereus and closely related low-GC Gram-positives. Environ Microbiol 2008;10:2796–2809 [CrossRef][PubMed]
    [Google Scholar]
  39. Francke C, Kerkhoven R, Wels M, Siezen RJ. A generic approach to identify Transcription Factor-specific operator motifs; inferences for LacI-family mediated regulation in Lactobacillus plantarum WCFS1. BMC Genomics 2008;9:145 [CrossRef][PubMed]
    [Google Scholar]
  40. Weickert MJ, Chambliss GH. Site-directed mutagenesis of a catabolite repression operator sequence in Bacillus subtilis. Proc Natl Acad Sci USA 1990;87:6238–6242 [CrossRef][PubMed]
    [Google Scholar]
  41. Kim HM, Park YH, Yoon CK, Seok YJ. Histidine phosphocarrier protein regulates pyruvate kinase A activity in response to glucose in Vibrio vulnificus. Mol Microbiol 2015;96:293–305 [CrossRef][PubMed]
    [Google Scholar]
  42. Charbonnier T, Le Coq D, McGovern S, Calabre M, Delumeau O et al. Molecular and physiological logics of the pyruvate-induced response of a novel transporter in Bacillus subtilis. MBio 2017;8: [CrossRef][PubMed]
    [Google Scholar]
  43. van den Esker MH, Kovács ÁT, Kuipers OP. YsbA and LytST are essential for pyruvate utilization in Bacillus subtilis. Environ Microbiol 2017;19:83–94 [CrossRef][PubMed]
    [Google Scholar]
  44. LeBlanc DJ, Lee LN, Abu-Al-Jaibat A. Molecular, genetic, and functional analysis of the basic replicon of pVA380-1, a plasmid of oral streptococcal origin. Plasmid 1992;28:130–145 [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000744
Loading
/content/journal/micro/10.1099/mic.0.000744
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