1887

Abstract

It is known that the (or ) gene encoding uridine 5′-(beta-1-threo-pentapyranosyl-4-ulose diphosphate) aminotransferase plays a critical role in colistin resistance in through the addition of 4-amino-4-deoxy--arabinose (-Ara4N) to lipid A. In this study, we attempted to obtain a colistin-resistant mutant from an -deleted mutant through exposure to colistin.

We constructed an deletion mutant (P5Δ:: ) from a colistin-susceptible strain (P5) by allelic replacement mutagenesis, and colistin-resistant mutants were selected using P5 and P5Δ:: . The growth rate, lipid A structure, biofilm-forming activity and cell viability in diverse stressful conditions (osmotic, oxidative, acidic and heat stress) were investigated. Expression of and was evaluated by qRT-PCR.

An deletion mutant was shown to develop colistin resistance through the addition of -Ara4N to lipid A, despite a low survival rate (over 1000-fold lower than that of the wild-type strain) in the media with colistin. Two colistin-resistant mutants showed higher survival rates than colistin-susceptible strains against 5 % NaCl. In the presence of acidic and heat stress, P5Δ:: -Cst exhibited higher survival rates during conditions of 1 % HCl and 42 °C than the other strains. Both and genes were overexpressed significantly in both colistin-resistant mutants, but and genes were not.

We revealed that colistin resistance could be developed despite deletion in through the addition of -Ara4N to lipid A, which was accompanied by diverse physiological changes.

Keyword(s): arnB , colistin and lipid A
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.000456
2017-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jmm/66/6/833.html?itemId=/content/journal/jmm/10.1099/jmm.0.000456&mimeType=html&fmt=ahah

References

  1. Lister PD, Wolter DJ, Hanson ND. Antibacterial-resistant Pseudomonas aeruginosa: clinical impact and complex regulation of chromosomally encoded resistance mechanisms. Clin Microbiol Rev 2009; 22:582–610 [View Article][PubMed]
    [Google Scholar]
  2. Hancock RE, Speert DP. Antibiotic resistance in Pseudomonas aeruginosa: mechanisms and impact on treatment. Drug Resist Updat 2000; 3:247–255 [View Article][PubMed]
    [Google Scholar]
  3. Nation RL, Li J. Colistin in the 21st century. Curr Opin Infect Dis 2009; 22:535–543 [View Article][PubMed]
    [Google Scholar]
  4. Evans ME, Feola DJ, Rapp RP. Polymyxin B sulfate and colistin: old antibiotics for emerging multiresistant gram-negative bacteria. Ann Pharmacother 1999; 33:960–967 [View Article][PubMed]
    [Google Scholar]
  5. Andersson DI, Hughes D, Kubicek-Sutherland JZ. Mechanisms and consequences of bacterial resistance to antimicrobial peptides. Drug Resist Updat 2016; 26:43–57 [View Article][PubMed]
    [Google Scholar]
  6. Landman D, Bratu S, Alam M, Quale J. Citywide emergence of Pseudomonas aeruginosa strains with reduced susceptibility to polymyxin B. J Antimicrob Chemother 2005; 55:954–957 [View Article][PubMed]
    [Google Scholar]
  7. Wang CY, Jerng JS, Chen KY, Cheng KY, Lee LN et al. Pandrug-resistant Pseudomonas aeruginosa among hospitalised patients: clinical features, risk-factors and outcomes. Clin Microbiol Infect 2006; 12:63–68 [View Article][PubMed]
    [Google Scholar]
  8. Lee JY, Song JH, Ko KS. Identification of nonclonal Pseudomonas aeruginosa isolates with reduced colistin susceptibility in Korea. Microb Drug Resist 2011; 17:299–304 [View Article][PubMed]
    [Google Scholar]
  9. Miller AK, Brannon MK, Stevens L, Johansen HK, Selgrade SE et al. PhoQ mutations promote lipid A modification and polymyxin resistance of Pseudomonas aeruginosa found in colistin-treated cystic fibrosis patients. Antimicrob Agents Chemother 2011; 55:5761–5769 [View Article][PubMed]
    [Google Scholar]
  10. Moskowitz SM, Ernst RK, Miller SI. PmrAB, a two-component regulatory system of Pseudomonas aeruginosa that modulates resistance to cationic antimicrobial peptides and addition of aminoarabinose to lipid A. J Bacteriol 2004; 186:575–579 [View Article][PubMed]
    [Google Scholar]
  11. Lee JY, Park YK, Chung ES, Na IY, Ko KS. Evolved resistance to colistin and its loss due to genetic reversion in Pseudomonas aeruginosa. Sci Rep 2016; 6:25543 [View Article][PubMed]
    [Google Scholar]
  12. Lee JY, Na IY, Park YK, Ko KS. Genomic variations between colistin-susceptible and -resistant Pseudomonas aeruginosa clinical isolates and their effects on colistin resistance. J Antimicrob Chemother 2014; 69:1248–1256 [View Article][PubMed]
    [Google Scholar]
  13. Riley M, Abe T, Arnaud MB, Berlyn MK, Blattner FR et al. Escherichia coli K-12: a cooperatively developed annotation snapshot—2005. Nucleic Acids Res 2006; 34:1–9 [View Article][PubMed]
    [Google Scholar]
  14. Mcphee JB, Bains M, Winsor G, Lewenza S, Kwasnicka A et al. Contribution of the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems to Mg2+-induced gene regulation in Pseudomonas aeruginosa. J Bacteriol 2006; 188:3995–4006 [View Article][PubMed]
    [Google Scholar]
  15. Schurek KN, Sampaio JL, Kiffer CR, Sinto S, Mendes CM et al. Involvement of pmrAB and phoPQ in polymyxin B adaptation and inducible resistance in non-cystic fibrosis clinical isolates of Pseudomonas aeruginosa. Antimicrob Agents Chemother 2009; 53:4345–4351 [View Article][PubMed]
    [Google Scholar]
  16. Fernández L, Gooderham WJ, Bains M, McPhee JB, Wiegand I et al. Adaptive resistance to the "last hope" antibiotics polymyxin B and colistin in Pseudomonas aeruginosa is mediated by the novel two-component regulatory system ParR-ParS. Antimicrob Agents Chemother 2010; 54:3372–3382 [View Article][PubMed]
    [Google Scholar]
  17. Muller C, Plésiat P, Jeannot K. A two-component regulatory system interconnects resistance to polymyxins, aminoglycosides, fluoroquinolones, and β-lactams in Pseudomonas aeruginosa. Antimicrob Agents Chemother 2011; 55:1211–1221 [View Article][PubMed]
    [Google Scholar]
  18. Fernández L, Jenssen H, Bains M, Wiegand I, Gooderham WJ et al. The two-component system CprRS senses cationic peptides and triggers adaptive resistance in Pseudomonas aeruginosa independently of ParRS. Antimicrob Agents Chemother 2012; 56:6212–6222 [View Article][PubMed]
    [Google Scholar]
  19. Lee JY, Chung ES, Na IY, Kim H, Shin D et al. Development of colistin resistance in pmrA-, phoP-, parR- and cprR-inactivated mutants of Pseudomonas aeruginosa. J Antimicrob Chemother 2014; 69:2966–2971 [View Article][PubMed]
    [Google Scholar]
  20. Song JH, Ko KS, Lee JY, Baek JY, Oh WS et al. Identification of essential genes in Streptococcus pneumoniae by allelic replacement mutagenesis. Mol Cells 2005; 19:365–374[PubMed]
    [Google Scholar]
  21. Schweizer HP. Escherichia-Pseudomonas shuttle vectors derived from pUC18/19. Gene 1991; 97:109–112 [View Article][PubMed]
    [Google Scholar]
  22. Datsenko KA, Wanner BL. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 2000; 97:6640–6645 [View Article][PubMed]
    [Google Scholar]
  23. Patel JB, Cockerill III FR, Alder J, Bradford PA, Eliopoulos GM et al. Performance Standards for Antimicrobial Susceptibility Testing; Twenty-Fourth Informational Supplement M100-S24 Wayne, PA: Clinical and Laboratory Standards Institute; 2014
    [Google Scholar]
  24. Pelletier MR, Casella LG, Jones JW, Adams MD, Zurawski DV et al. Unique structural modifications are present in the lipopolysaccharide from colistin-resistant strains of Acinetobacter baumannii. Antimicrob Agents Chemother 2013; 57:4831–4840 [View Article][PubMed]
    [Google Scholar]
  25. Ernst RK, Yi EC, Guo L, Lim KB, Burns JL et al. Specific lipopolysaccharide found in cystic fibrosis airway Pseudomonas aeruginosa. Science 1999; 286:1561–1565 [View Article][PubMed]
    [Google Scholar]
  26. Park YK, Lee JY, Ko KS. Transcriptomic analysis of colistin-susceptible and colistin-resistant isolates identifies genes associated with colistin resistance in Acinetobacter baumannii. Clin Microbiol Infect 2015; 21:765.e1–765.e7 [View Article][PubMed]
    [Google Scholar]
  27. Merritt JH, Kadouri DE, O'Toole GA. Growing and analyzing static biofilms. Curr Protoc Microbiol 2005; Chapter 1:1:1B.1–1B.1.17 [View Article][PubMed]
    [Google Scholar]
  28. Wang X, Kim Y, Ma Q, Hong SH, Pokusaeva K et al. Cryptic prophages help bacteria cope with adverse environments. Nat Commun 2010; 1:147 [View Article][PubMed]
    [Google Scholar]
  29. Park YK, Choi JY, Shin D, Ko KS. Correlation between overexpression and amino acid substitution of the PmrAB locus and colistin resistance in Acinetobacter baumannii. Int J Antimicrob Agents 2011; 37:525–530 [View Article][PubMed]
    [Google Scholar]
  30. Choi MJ, Kim S, Ko KS. Pathways regulating the pbgP operon and colistin resistance in Klebsiella pneumoniae strains. J Microbiol Biotechnol 2016; 26:1620–1628 [View Article][PubMed]
    [Google Scholar]
  31. Liu YY, Wang Y, Walsh TR, Yi LX, Zhang R et al. Emergence of plasmid-mediated colistin resistance mechanism MCR-1 in animals and human beings in China: a microbiological and molecular biological study. Lancet Infect Dis 2016; 16:161–168 [View Article][PubMed]
    [Google Scholar]
  32. Bader MW, Sanowar S, Daley ME, Schneider AR, Cho U et al. Recognition of antimicrobial peptides by a bacterial sensor kinase. Cell 2005; 122:461–472 [View Article][PubMed]
    [Google Scholar]
  33. Tam C, Missiakas D. Changes in lipopolysaccharide structure induce the σE-dependent response of Escherichia coli. Mol Microbiol 2005; 55:1403–1412 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.000456
Loading
/content/journal/jmm/10.1099/jmm.0.000456
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