1887

Abstract

in dental biofilms often faces life-threatening threats such as killing by antimicrobial molecules from competing species or from the host. The ability of to cope with such threats is crucial for its survival and persistence in dental biofilms. By screening a transposon mutant library, we identified 11 transposon insertion mutants that were sensitive to bacitracin. Two of these mutants, XTn-01 and XTn-03, had an independent insertion in the same locus, SMU.244, which encoded a homologue of undecaprenyl pyrophosphate phosphatase (UppP). In this study, we describe the genetic and phenotypic characterization of SMU.244 in antibiotic resistance. The results revealed that deletion of SMU.244 results in a mutant (XTΔ244) that is highly sensitive to bacitracin, but confers more resistance to lactococcin G, a class IIb bacteriocin. Introduction of the intact SMU.244 into XTΔ244 completely restores its resistance to bacitracin and the susceptibility to lactococcin G. The XTΔ244 was also defective in forming the WT biofilm, although its growth was not significantly affected. Using recombinant protein technology, we demonstrated that the SMU.244-encoded protein displays enzyme activity to catalyse dephosphorylation of the substrate. The transcriptional reporter assays showed that maintains a moderate level of expression of SMU.244 in the absence of bacitracin, but bacitracin at sub-MICs can further induce its expression. We concluded that SMU.244 encodes an UppP protein that plays important roles in cell wall biosynthesis and bacitracin resistance in . The results described here may further our understanding of the molecular mechanisms by which copes with antibiotics such as bacitracin.

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.000142
2015-09-01
2024-04-20
Loading full text...

Full text loading...

/deliver/fulltext/micro/161/9/1857.html?itemId=/content/journal/micro/10.1099/mic.0.000142&mimeType=html&fmt=ahah

References

  1. Ajdić D., McShan W. M., McLaughlin R. E., Savić G., Chang J., Carson M. B., Primeaux C., Tian R., Kenton S., other authors. 2002; Genome sequence of Streptococcus mutans UA159, a cariogenic dental pathogen. Proc Natl Acad Sci U S A 99:14434–14439 [View Article][PubMed]
    [Google Scholar]
  2. Bernard R., El Ghachi M., Mengin-Lecreulx D., Chippaux M., Denizot F. 2005; BcrC from Bacillus subtilis acts as an undecaprenyl pyrophosphate phosphatase in bacitracin resistance. J Biol Chem 280:28852–28857 [View Article][PubMed]
    [Google Scholar]
  3. Bickford J. S., Nick H. S. 2013; Conservation of the PTEN catalytic motif in the bacterial undecaprenyl pyrophosphate phosphatase. BacA/UppP. Microbiology 159:2444–2455 [View Article][PubMed]
    [Google Scholar]
  4. Chalker A. F., Ingraham K. A., Lunsford R. D., Bryant A. P., Bryant J., Wallis N. G., Broskey J. P., Pearson S. C., Holmes D. J. 2000; The bacA gene, which determines bacitracin susceptibility in Streptococcus pneumoniae and Staphylococcus aureus, is also required for virulence. Microbiology 146:1547–1553[PubMed] [CrossRef]
    [Google Scholar]
  5. Chang H. Y., Chou C. C., Hsu M. F., Wang A. H. 2014; Proposed carrier lipid-binding site of undecaprenyl pyrophosphate phosphatase from Escherichia coli . J Biol Chem 289:18719–18735 [View Article][PubMed]
    [Google Scholar]
  6. Chen Q., Wu H., Fives-Taylor P. M. 2002; Construction of a novel transposon mutagenesis system useful in the isolation of Streptococcus parasanguis mutants defective in Fap1 glycosylation. Infect Immun 70:6534–6540 [View Article][PubMed]
    [Google Scholar]
  7. Cotter P. D. 2014; An ‘Upp’-turn in bacteriocin receptor identification. Mol Microbiol 92:1159–1163 [View Article][PubMed]
    [Google Scholar]
  8. Dale B. A., Fredericks L. P. 2005; Antimicrobial peptides in the oral environment: expression and function in health and disease. Curr Issues Mol Biol 7:119–133[PubMed]
    [Google Scholar]
  9. Desai K., Mashburn-Warren L., Federle M. J., Morrison D. A. 2012; Development of competence for genetic transformation of Streptococcus mutans in a chemically defined medium. J Bacteriol 194:3774–3780 [View Article][PubMed]
    [Google Scholar]
  10. Diamond G., Beckloff N., Ryan L. K. 2008; Host defense peptides in the oral cavity and the lung: similarities and differences. J Dent Res 87:915–927 [View Article][PubMed]
    [Google Scholar]
  11. Dong G., Tian X. L., Gomez Z. A., Li Y. H. 2014; Regulated proteolysis of the alternative sigma factor SigX in Streptococcus mutans: implication in the escape from competence. BMC Microbiol 14:183 [View Article][PubMed]
    [Google Scholar]
  12. El Ghachi M., Bouhss A., Blanot D., Mengin-Lecreulx D. 2004; The bacA gene of Escherichia coli encodes an undecaprenyl pyrophosphate phosphatase activity. J Biol Chem 279:30106–30113 [View Article][PubMed]
    [Google Scholar]
  13. El Ghachi M., Derbise A., Bouhss A., Mengin-Lecreulx D. 2005; Identification of multiple genes encoding membrane proteins with undecaprenyl pyrophosphate phosphatase (UppP) activity in Escherichia coli . J Biol Chem 280:18689–18695 [View Article][PubMed]
    [Google Scholar]
  14. Gong Y., Tian X. L., Sutherland T., Sisson G., Mai J., Ling J., Li Y. H. 2009; Global transcriptional analysis of acid-inducible genes in Streptococcus mutans: multiple two-component systems involved in acid adaptation. Microbiology 155:3322–3332 [View Article][PubMed]
    [Google Scholar]
  15. Hsu M. F., Yu T. F., Chou C. C., Fu H. Y., Yang C. S., Wang A. H. J. 2013; Using Haloarcula marismortui bacteriorhodopsin as a fusion tag for enhancing and visible expression of integral membrane proteins in Escherichia coli . PLoS One 8:e56363 [View Article][PubMed]
    [Google Scholar]
  16. Jordan S., Hutchings M. I., Mascher T. 2008; Cell envelope stress response in Gram-positive bacteria. FEMS Microbiol Rev 32:107–146 [View Article][PubMed]
    [Google Scholar]
  17. Karlyshev A. V., Pallen M. J., Wren B. W. 2000; Single-primer PCR procedure for rapid identification of transposon insertion sites. Biotechniques 1080:1082[PubMed]
    [Google Scholar]
  18. Kim J. K., Lee H. J., Kikuchi Y., Kitagawa W., Nikoh N., Fukatsu T., Lee B. L. 2013; Bacterial cell wall synthesis gene uppP is required for Burkholderia colonization of the stinkbug gut. Appl Environ Microbiol 79:4879–4886 [View Article][PubMed]
    [Google Scholar]
  19. Kjos M., Oppegård C., Diep D. B., Nes I. F., Veening J. W., Nissen-Meyer J., Kristensen T. 2014; Sensitivity to the two-peptide bacteriocin lactococcin G is dependent on UppP, an enzyme involved in cell-wall synthesis. Mol Microbiol 92:1177–1187 [View Article][PubMed]
    [Google Scholar]
  20. Kreth J., Zhang Y., Herzberg M. C. 2008; Streptococcal antagonism in oral biofilms: Streptococcus sanguinis and Streptococcus gordonii interference with Streptococcus mutans . J Bacteriol 190:4632–4640 [View Article][PubMed]
    [Google Scholar]
  21. Kuramitsu H. K., He X., Lux R., Anderson M. H., Shi W. 2007; Interspecies interactions within oral microbial communities. Microbiol Mol Biol Rev 71:653–670 [View Article][PubMed]
    [Google Scholar]
  22. Lau P. C. Y., Sung C. K., Lee J. H., Morrison D. A., Cvitkovitch D. G. 2002; PCR ligation mutagenesis in transformable streptococci: application and efficiency. J Microbiol Methods 49:193–205 [View Article][PubMed]
    [Google Scholar]
  23. LeBlanc D. J., Lee L. N., Abu-Al-Jaibat A. 1992; Molecular, genetic, and functional analysis of the basic replicon of pVA380-1, a plasmid of oral streptococcal origin. Plasmid 28:130–145 [View Article][PubMed]
    [Google Scholar]
  24. Li Y. H., Tian X. 2012; Quorum sensing and bacterial social interactions in biofilms. Sensors (Basel) 12:2519–2538 [View Article][PubMed]
    [Google Scholar]
  25. Li Y. H., Tang N., Aspiras M. B., Lau P. C., Lee J. H., Ellen R. P., Cvitkovitch D. G. 2002; A quorum-sensing signaling system essential for genetic competence in Streptococcus mutans is involved in biofilm formation. J Bacteriol 184:2699–2708 [View Article][PubMed]
    [Google Scholar]
  26. Li Y. H., Tian X. L., Layton G., Norgaard C., Sisson G. 2008; Additive attenuation of virulence and cariogenic potential of Streptococcus mutans by simultaneous inactivation of the ComCDE quorum-sensing system and HK/RR11 two-component regulatory system. Microbiology 154:3256–3265 [View Article][PubMed]
    [Google Scholar]
  27. Liljemark W. F., Okrent D. H., Bloomquist C. G. 1976; Differential recovery of Streptococcus mutans from various mitis-salivarius agar preparations. J Clin Microbiol 4:108–109[PubMed]
    [Google Scholar]
  28. Lis M., Kuramitsu H. K. 2003; The stress-responsive dgk gene from Streptococcus mutans encodes a putative undecaprenol kinase activity. Infect Immun 71:1938–1943 [View Article][PubMed]
    [Google Scholar]
  29. Loo C. Y., Corliss D. A., Ganeshkumar N. 2000; Streptococcus gordonii biofilm formation: identification of genes that code for biofilm phenotypes. J Bacteriol 182:1374–1382 [View Article][PubMed]
    [Google Scholar]
  30. Mai J., Tian X. L., Gallant J. W., Merkley N., Biswas Z., Syvitski R., Douglas S. E., Ling J., Li Y. H. 2011; A novel target-specific, salt-resistant antimicrobial peptide against the cariogenic pathogen Streptococcus mutans . Antimicrob Agents Chemother 55:5205–5213 [View Article][PubMed]
    [Google Scholar]
  31. Mascher T., Helmann J. D., Unden G. 2006; Stimulus perception in bacterial signal-transducing histidine kinases. Microbiol Mol Biol Rev 70:910–938 [View Article][PubMed]
    [Google Scholar]
  32. Mesak L. R., Yim G., Davies J. 2009; Improved lux reporters for use in Staphylococcus aureus . Plasmid 61:182–187 [View Article][PubMed]
    [Google Scholar]
  33. Oppegård C., Fimland G., Thorbaek L., Nissen-Meyer J. 2007; Analysis of the two-peptide bacteriocins lactococcin G and enterocin 1071 by site-directed mutagenesis. Appl Environ Microbiol 73:2931–2938 [View Article][PubMed]
    [Google Scholar]
  34. Ouyang J., Tian X. L., Versey J., Wishart A., Li Y. H. 2010; The BceABRS four-component system regulates the bacitracin-induced cell envelope stress response in Streptococcus mutans . Antimicrob Agents Chemother 54:3895–3906 [View Article][PubMed]
    [Google Scholar]
  35. Subrt N., Mesak L. R., Davies J. 2011; Modulation of virulence gene expression by cell wall active antibiotics in Staphylococcus aureus . J Antimicrob Chemother 66:979–984 [View Article][PubMed]
    [Google Scholar]
  36. Suntharalingam P., Senadheera M. D., Mair R. W., Lévesque C. M., Cvitkovitch D. G. 2009; The LiaFSR system regulates the cell envelope stress response in Streptococcus mutans . J Bacteriol 191:2973–2984 [View Article][PubMed]
    [Google Scholar]
  37. Tatar L. D., Marolda C. L., Polischuk A. N., van Leeuwen D., Valvano M. A. 2007; An Escherichia coli undecaprenyl-pyrophosphate phosphatase implicated in undecaprenyl phosphate recycling. Microbiology 153:2518–2529 [View Article][PubMed]
    [Google Scholar]
  38. Tian X. L., Dong G., Liu T., Gomez Z. A., Wahl A., Hols P., Li Y. H. 2013; MecA protein acts as a negative regulator of genetic competence in Streptococcus mutans . J Bacteriol 195:5196–5206 [View Article][PubMed]
    [Google Scholar]
  39. Tsuda H., Yamashita Y., Shibata Y., Nakano Y., Koga T. 2002; Genes involved in bacitracin resistance in Streptococcus mutans . Antimicrob Agents Chemother 46:3756–3764 [View Article][PubMed]
    [Google Scholar]
  40. Yamashita Y., Tsukioka Y., Tomihisa K., Nakano Y., Koga T. 1998; Genes involved in cell wall localization and side chain formation of rhamnose-glucose polysaccharide in Streptococcus mutans . J Bacteriol 180:5803–5807[PubMed]
    [Google Scholar]
  41. Yamashita Y., Shibata Y., Nakano Y., Tsuda H., Kido N., Ohta M., Koga T. 1999; A novel gene required for rhamnose-glucose polysaccharide synthesis in Streptococcus mutans . J Bacteriol 181:6556–6559[PubMed]
    [Google Scholar]
  42. Zhang J., Biswas I. 2009; 3′-Phosphoadenosine-5′-phosphate phosphatase activity is required for superoxide stress tolerance in Streptococcus mutans . J Bacteriol 191:4330–4340 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.000142
Loading
/content/journal/micro/10.1099/mic.0.000142
Loading

Data & Media loading...

Supplements

Supplementary Data

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