1887

Abstract

The type strain ATCC 14579 harbours pBClin15, a linear plasmid with similar genome organization to tectiviruses. Since phage morphogenesis is not known to occur it has been suggested that pBClin15 may be a defect relic of a tectiviral prophage without relevance for the bacterial physiology. However, in this paper, we demonstrate that a pBClin15-cured strain is more tolerant to antibiotics interfering with DNA integrity than the WT strain. Growth in the presence of crystal violet or the quinolones nalidixic acid, norfloxacin or ciprofloxacin resulted in aggregation and lysis of the WT strain, whereas the pBClin15-cured strain was unaffected. Microarray analysis comparing the gene expression in the WT and pBClin15-cured strains showed that pBClin15 gene expression was strongly upregulated in response to norfloxacin stress, and coincided with lysis and aggregation of the WT strain. The aggregating bacteria experienced a significant survival benefit compared with the planktonic counterparts in the presence of norfloxacin. There was no difference between the WT and pBClin15-cured strains during growth in the absence of norfloxacin, the pBClin15 genes were moderately expressed, and no effect was observed on chromosomal gene expression. These data demonstrate for the first time that although pBClin15 may be a remnant of a temperate phage, it negatively affects the DNA stress tolerance of ATCC 14579. Furthermore, our results warrant a recommendation to always verify the presence of pBClin15 following genetic manipulation of ATCC 14579.

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2013-11-01
2024-03-28
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References

  1. Ackermann H. W., Roy R., Martin M., Murthy M. R., Smirnoff W. A.( 1978). Partial characterization of a cubic Bacillus phage. Can J Microbiol 24:986–993 [View Article][PubMed]
    [Google Scholar]
  2. Balaban N. Q., Merrin J., Chait R., Kowalik L., Leibler S.( 2004). Bacterial persistence as a phenotypic switch. Science 305:1622–1625 [View Article][PubMed]
    [Google Scholar]
  3. Beloin C., Valle J., Latour-Lambert P., Faure P., Kzreminski M., Balestrino D., Haagensen J. A., Molin S., Prensier G. et al.( 2004). Global impact of mature biofilm lifestyle on Escherichia coli K-12 gene expression. Mol Microbiol 51:659–674 [View Article][PubMed]
    [Google Scholar]
  4. Bishop-Lilly K. A., Plaut R. D., Chen P. E., Akmal A., Willner K. M., Butani A., Dorsey S., Mokashi V., Mateczun A. J. et al.( 2012). Whole genome sequencing of phage resistant Bacillus anthracis mutants reveals an essential role for cell surface anchoring protein CsaB in phage AP50c adsorption. Virol J 9:246 [View Article][PubMed]
    [Google Scholar]
  5. Brissette J. L., Russel M., Weiner L., Model P.( 1990). Phage shock protein, a stress protein of Escherichia coli. Proc Natl Acad Sci U S A 87:862–866 [View Article][PubMed]
    [Google Scholar]
  6. Butala M., Zgur-Bertok D., Busby S. J.( 2009). The bacterial LexA transcriptional repressor. Cell Mol Life Sci 66:82–93 [View Article][PubMed]
    [Google Scholar]
  7. Carlson C. R., Johansen T., Kolstø A. B.( 1996). The chromosome map of Bacillus thuringiensis subsp. canadensis HD224 is highly similar to that of the Bacillus cereus type strain ATCC 14579. FEMS Microbiol Lett 141:163–167 [View Article][PubMed]
    [Google Scholar]
  8. Chai Y., Chu F., Kolter R., Losick R.( 2008). Bistability and biofilm formation in Bacillus subtilis. Mol Microbiol 67:254–263 [View Article][PubMed]
    [Google Scholar]
  9. Champoux J. J.( 2001). DNA topoisomerases: structure, function, and mechanism. Annu Rev Biochem 70:369–413 [View Article][PubMed]
    [Google Scholar]
  10. Darwin A. J.( 2005). The phage-shock-protein response. Mol Microbiol 57:621–628 [View Article][PubMed]
    [Google Scholar]
  11. Diver J. M.( 1989). Quinolone uptake by bacteria and bacterial killing. Rev Infect Dis 11:Suppl. 5S941–S946 [View Article][PubMed]
    [Google Scholar]
  12. Docampo R., Moreno S. N.( 1990). The metabolism and mode of action of gentian violet. Drug Metab Rev 22:161–178 [View Article][PubMed]
    [Google Scholar]
  13. Dörr T., Lewis K., Vulić M.( 2009). SOS response induces persistence to fluoroquinolones in Escherichia coli. PLoS Genet 5:e1000760 [View Article][PubMed]
    [Google Scholar]
  14. Drlica K.( 1999). Mechanism of fluoroquinolone action. Curr Opin Microbiol 2:504–508 [View Article][PubMed]
    [Google Scholar]
  15. Drlica K., Malik M., Kerns R. J., Zhao X.( 2008). Quinolone-mediated bacterial death. Antimicrob Agents Chemother 52:385–392 [View Article][PubMed]
    [Google Scholar]
  16. Drlica K., Hiasa H., Kerns R., Malik M., Mustaev A., Zhao X.( 2009). Quinolones: action and resistance updated. Curr Top Med Chem 9:981–998 [View Article][PubMed]
    [Google Scholar]
  17. Fornelos N., Bamford J. K., Mahillon J.( 2011). Phage-borne factors and host LexA regulate the lytic switch in phage GIL01. J Bacteriol 193:6008–6019 [View Article][PubMed]
    [Google Scholar]
  18. Gentleman R. C., Carey V. J., Bates D. M., Bolstad B., Dettling M., Dudoit S., Ellis B., Gautier L., Ge Y. et al.( 2004). Bioconductor: open software development for computational biology and bioinformatics. Genome Biol 5:R80 [View Article][PubMed]
    [Google Scholar]
  19. Gohar M., Faegri K., Perchat S., Ravnum S., Økstad O. A., Gominet M., Kolstø A. B., Lereclus D.( 2008). The PlcR virulence regulon of Bacillus cereus.. PLoS ONE 3:e2793 [View Article][PubMed]
    [Google Scholar]
  20. Ivanova N., Sorokin A., Anderson I., Galleron N., Candelon B., Kapatral V., Bhattacharyya A., Reznik G., Mikhailova N. et al.( 2003). Genome sequence of Bacillus cereus and comparative analysis with Bacillus anthracis. Nature 423:87–91 [View Article][PubMed]
    [Google Scholar]
  21. Janes B. K., Stibitz S.( 2006). Routine markerless gene replacement in Bacillus anthracis. Infect Immun 74:1949–1953 [View Article][PubMed]
    [Google Scholar]
  22. Kearns D. B., Chu F., Rudner R., Losick R.( 2004). Genes governing swarming in Bacillus subtilis and evidence for a phase variation mechanism controlling surface motility. Mol Microbiol 52:357–369 [View Article][PubMed]
    [Google Scholar]
  23. Kotiranta A., Haapasalo M., Kari K., Kerosuo E., Olsen I., Sorsa T., Meurman J. H., Lounatmaa K.( 1998). Surface structure, hydrophobicity, phagocytosis, and adherence to matrix proteins of Bacillus cereus cells with and without the crystalline surface protein layer. Infect Immun 66:4895–4902[PubMed]
    [Google Scholar]
  24. Lewis K.( 2007). Persister cells, dormancy and infectious disease. Nat Rev Microbiol 5:48–56 [View Article][PubMed]
    [Google Scholar]
  25. Lopez D., Vlamakis H., Kolter R.( 2009). Generation of multiple cell types in Bacillus subtilis. FEMS Microbiol Rev 33:152–163 [View Article][PubMed]
    [Google Scholar]
  26. Love P. E., Yasbin R. E.( 1984). Genetic characterization of the inducible SOS-like system of Bacillus subtilis. J Bacteriol 160:910–920[PubMed]
    [Google Scholar]
  27. Maamar H., Dubnau D.( 2005). Bistability in the Bacillus subtilis K-state (competence) system requires a positive feedback loop. Mol Microbiol 56:615–624 [View Article][PubMed]
    [Google Scholar]
  28. Mignot T., Denis B., Couture-Tosi E., Kolstø A. B., Mock M., Fouet A.( 2001). Distribution of S-layers on the surface of Bacillus cereus strains: phylogenetic origin and ecological pressure. Environ Microbiol 3:493–501 [View Article][PubMed]
    [Google Scholar]
  29. Nagy E.( 1974). A highly specific phage attacking Bacillus anthracis strain Sterne. Acta Microbiol Acad Sci Hung 21:257–263[PubMed]
    [Google Scholar]
  30. Nagy E., Ivánovics G.( 1977). Association of probable defective phage particles with lysis by bacteriophage AP50 in Bacillus anthracis. J Gen Microbiol 102:215–219 [View Article][PubMed]
    [Google Scholar]
  31. O’Toole G., Kaplan H. B., Kolter R.( 2000). Biofilm formation as microbial development. Annu Rev Microbiol 54:49–79 [View Article][PubMed]
    [Google Scholar]
  32. Olsen R. H., Siak J. S., Gray R. H.( 1974). Characteristics of PRD1, a plasmid-dependent broad host range DNA bacteriophage. J Virol 14:689–699[PubMed]
    [Google Scholar]
  33. Shah D., Zhang Z., Khodursky A., Kaldalu N., Kurg K., Lewis K.( 2006). Persisters: a distinct physiological state of E. coli. BMC Microbiol 6:53 [View Article][PubMed]
    [Google Scholar]
  34. Simmons L. A., Goranov A. I., Kobayashi H., Davies B. W., Yuan D. S., Grossman A. D., Walker G. C.( 2009). Comparison of responses to double-strand breaks between Escherichia coli and Bacillus subtilis reveals different requirements for SOS induction. J Bacteriol 191:1152–1161 [View Article][PubMed]
    [Google Scholar]
  35. Smyth G. K.( 2005). Limma: linear models for microarray data. Bioinformatics and Computational Biology Solutions Using R and Bioconductor397–420 Gentleman V. C. R., Dudoit S., Irizarry R., Huber W. Berlin: Springer; [View Article]
    [Google Scholar]
  36. Sozhamannan S., McKinstry M., Lentz S. M., Jalasvuori M., McAfee F., Smith A., Dabbs J., Ackermann H. W., Bamford J. K. et al.( 2008). Molecular characterization of a variant of Bacillus anthracis-specific phage AP50 with improved bacteriolytic activity. Appl Environ Microbiol 74:6792–6796 [View Article][PubMed]
    [Google Scholar]
  37. Stabell F. B., Egge-Jacobsen W., Risøen P. A., Kolstø A. B., Økstad O. A.( 2009). ORF 2 from the Bacillus cereus linear plasmid pBClin15 encodes a DNA binding protein. Lett Appl Microbiol 48:51–57 [View Article][PubMed]
    [Google Scholar]
  38. Strömsten N. J., Benson S. D., Burnett R. M., Bamford D. H., Bamford J. K.( 2003). The Bacillus thuringiensis linear double-stranded DNA phage Bam35, which is highly similar to the Bacillus cereus linear plasmid pBClin15, has a prophage state. J Bacteriol 185:6985–6989 [View Article][PubMed]
    [Google Scholar]
  39. Veening J. W., Hamoen L. W., Kuipers O. P.( 2005). Phosphatases modulate the bistable sporulation gene expression pattern in Bacillus subtilis. Mol Microbiol 56:1481–1494 [View Article][PubMed]
    [Google Scholar]
  40. Verheust C., Jensen G., Mahillon J.( 2003). pGIL01, a linear tectiviral plasmid prophage originating from Bacillus thuringiensis serovar israelensis. Microbiology 149:2083–2092 [View Article][PubMed]
    [Google Scholar]
  41. Verheust C., Fornelos N., Mahillon J.( 2004). The Bacillus thuringiensis phage GIL01 encodes two enzymes with peptidoglycan hydrolase activity. FEMS Microbiol Lett 237:289–295[PubMed]
    [Google Scholar]
  42. Verheust C., Fornelos N., Mahillon J.( 2005). GIL16, a new gram-positive tectiviral phage related to the Bacillus thuringiensis GIL01 and the Bacillus cereus pBClin15 elements. J Bacteriol 187:1966–1973 [View Article][PubMed]
    [Google Scholar]
  43. Vilain S., Pretorius J. M., Theron J., Brözel V. S.( 2009). DNA as an adhesin: Bacillus cereus requires extracellular DNA to form biofilms. Appl Environ Microbiol 75:2861–2868 [View Article][PubMed]
    [Google Scholar]
  44. Wakelin L. P., Adams A., Hunter C., Waring M. J.( 1981). Interaction of crystal violet with nucleic acids. Biochemistry 20:5779–5787 [View Article][PubMed]
    [Google Scholar]
  45. Wolfe A. D.( 1977). Influence of cationic triphenylmethane dyes upon DNA polymerization and product hydrolysis by Escherichia coli polymerase I. Biochemistry 16:30–33 [View Article][PubMed]
    [Google Scholar]
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