1887

Microbiology Society logo

Volume 152, Issue 3

Phosphoethanolamine substitution in the lipid A of O157 : H7 and its association with PmrC Free

Abstract

This study shows that lipid A of O157 : H7 differs from that of K-12 in that it has a phosphoform at the C-1 position, which is distinctively modified by a phosphoethanolamine (PEtN) moiety, in addition to the diphosphoryl form. The gene responsible for the addition of PEtN to the lipid A of O157 : H7 was inactivated and the changes in lipid A profiles were assessed. The null mutant still produced PEtN-modified lipid A species, albeit in a reduced amount, indicating that PmrC was not the only enzyme that could be used to add PEtN to lipid A. Natural PEtN substitution was shown to be present in the lipid A of other serotypes of enterohaemorrhagic and absent from the lipid A of K-12. However, the cloned gene in a high-copy-number plasmid generated a large amount of PEtN-substituted lipid A species in K-12. The occurrence of PEtN-substituted lipid A species was associated with a slight increase in the MICs of cationic peptide antibiotics, suggesting that the lipid A modification with PEtN would be beneficial for survival of O157 : H7 in certain environmental niches. However, PEtN substitution in the lipid A profiles was not detected when putative inner-membrane proteins (YhbX/YbiP/YijP/Ecf3) that show significant similarity with PmrC in amino acid sequence were expressed from high-copy-number plasmids in K-12. This suggests that these potential homologues are not responsible for the addition of PEtN to lipid A in the mutant of O157 : H7. When cells were treated with EDTA, the amount of palmitoylated lipid A from the cells carrying a high-copy-number plasmid clone of that resulted in significant increase of PEtN substitution was unchanged compared with cells without PEtN substitution, suggesting that the PEtN moiety substituted in lipid A does not compensate for the loss of divalent cations required for bridging neighbouring lipid A molecules.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): CAMP, cationic antimicrobial peptide , CM, cytoplasmic membrane , EHEC, enterohaemorrhagic E. coli , l-Ara4N, 4-amino-4-deoxy-l-arabinose , MALDI-TOF, matrix-assisted laser desorption-ionization/time of flight , OM, outer membrane , PEtN, phosphoethanolamine , PMB, polymyxin B , PMBN, polymyxin B nonapeptide , STEC, Shiga toxin-producing E. coli and TM, transmembrane domain
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.28692-0
2006-03-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/152/3/657.html?itemId=/content/journal/micro/10.1099/mic.0.28692-0&mimeType=html&fmt=ahah

References

  1. Bishop R. E. 2005; The lipid A palmitoyltransferase PagP: molecular mechanisms and role in bacterial pathogenesis. Mol Microbiol 57:900–912 [CrossRef]
     [Google Scholar]
  2. Bligh E. G, Dyer W. J. 1959; A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917 [CrossRef]
     [Google Scholar]
  3. Boerlin P, Chen S, Colbourne J. K, Johnson R, De Grandis S, Gyles C. L. 1998; Evolution of enterohemorrhagic Escherichia coli hemolysin plasmids and the locus for enterocyte effacement in Shiga toxin-producing E. coli . Infect Immun 66:2553–2561
     [Google Scholar]
  4. Cox A. D, Wright J. C, Li J, Hood D. W, Moxon E. R, Richards J. C. 2003; Phosphorylation of the lipid A region of meningococcal lipopolysaccharide: identification of a family of transferases that add phosphoethanolamine to lipopolysaccharide. J Bacteriol 185:3270–3277 [CrossRef]
     [Google Scholar]
  5. Edwards R. A, Keller L. H, Schifferli D. M. 1998; Improved allelic exchange vectors and their use to analyze 987P fimbria gene expression. Gene 207:149–157 [CrossRef]
     [Google Scholar]
  6. Gibbons H. S, Kalb S. R, Cotter R. J, Raetz C. R. 2005; Role of Mg[sup]2+[/sup] and pH in the modification of Salmonella lipid A after endocytosis by macrophage tumour cells. Mol Microbiol 55:425–440
     [Google Scholar]
  7. Groisman E. A. 2001; The pleiotropic two-component regulatory system PhoP-PhoQ. J Bacteriol 183:1835–1842 [CrossRef]
     [Google Scholar]
  8. Groisman E. A, Kayser J, Soncini F. C. 1997; Regulation of polymyxin resistance and adaptation to low-Mg[sup]2+[/sup] environments. J Bacteriol 179:7040–7045
     [Google Scholar]
  9. Gunn J. S, Miller S. I. 1996; PhoP-PhoQ activates transcription of pmrAB , encoding a two-component regulatory system involved in Salmonella typhimurium antimicrobial peptide resistance. J Bacteriol 178:6857–6864
     [Google Scholar]
  10. Gunn J. S, Lim K. B, Krueger J, Kim K, Guo L, Hackett M, Miller S. I. 1998; PmrA-PmrB-regulated genes necessary for 4-aminoarabinose lipid A modification and polymyxin resistance. Mol Microbiol 27:1171–1182 [CrossRef]
     [Google Scholar]
  11. Hancock R. E. W. 1997; The bacterial outer membrane as a drug barrier. Trends Microbiol 5:37–42 [CrossRef]
     [Google Scholar]
  12. Holst O, Ulmer A. J, Brade H, Flad H. D, Rietschel E. T. 1996; Biochemistry and cell biology of bacterial endotoxins. FEMS Immunol Med Microbiol 16:83–104 [CrossRef]
     [Google Scholar]
  13. Jia W, Zoeiby A. E, Petruzziello T. N, Jayabalasingham B, Seyedirashti S, Bishop R. E. 2004; Lipid trafficking controls endotoxin acylation in outer membranes of Escherichia coli . J Biol Chem 279:44966–44975 [CrossRef]
     [Google Scholar]
  14. Kaniuk N. A, Vinogradov E, Li J, Monteiro M. A, Whitfield C. 2004; Chromosomal and plasmid-encoded enzymes are required for assembly of the R3-type core oligosaccharide in the lipopolysaccharide of Escherichia coli O157 : H7. J Biol Chem 279:31237–31250 [CrossRef]
     [Google Scholar]
  15. Kim S.-H, Jia W, Bishop R. E, Gyles C. 2004; An msbB homologue carried in plasmid pO157 encodes an acyltransferase involved in lipid A biosynthesis in Escherichia coli O157 : H7. Infect Immun 72:1174–1180 [CrossRef]
     [Google Scholar]
  16. Lee H, Hsu F. F, Turk J, Groisman E. A. 2004; The PmrA-regulated pmrC gene mediates phosphoethanolamine modification of lipid A and polymyxin resistance in Salmonella enterica . J Bacteriol 186:4124–4133 [CrossRef]
     [Google Scholar]
  17. Menard R, Sansonetti P. J, Parsot C. 1993; Nonpolar mutagenesis of the ipa genes defines IpaB, IpaC, and IpaD as effectors of Shigella flexneri entry into epithelial cells. J Bacteriol 175:5899–5906
     [Google Scholar]
  18. Miller J. H. 1992 A Short Course in Bacterial Genetics: a Laboratory Manual and Handbook for Escherichia coli and Related Bacteria Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  19. NCCLS 2002; Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals: approved standard. , 2nd edn.. 22M31–MA2 Wayne, PA: NCCLS;
  20. Nikaido H. 2003; Molecular basis of bacterial outer membrane permeability revisited. Microbiol Mol Biol Rev 67:593–656 [CrossRef]
     [Google Scholar]
  21. Nikaido H, Vaara M. 1985; Molecular basis of bacterial outer membrane permeability. Microbiol Rev 49:1–32
     [Google Scholar]
  22. Nummila K, Kilpelainen I, Zahringer U, Vaara M, Helander I. M. 1995; Lipopolysaccharides of polymyxin B-resistant mutants of Escherichia coli are extensively substituted by 2-aminoethyl pyrophosphate and contain aminoarabinose in lipid A. Mol Microbiol 16:271–278 [CrossRef]
     [Google Scholar]
  23. Raetz C. R. H. 2001; Regulated covalent modifications of lipid A. J Endotoxin Res 7:73–78 [CrossRef]
     [Google Scholar]
  24. Raetz C. R. H, Whitfield C. 2002; Lipopolysaccharide endotoxins. Annu Rev Biochem 71:635–700 [CrossRef]
     [Google Scholar]
  25. Reynolds C. M, Kalb S. R, Cotter R. J, Raetz C. R. 2005; A phosphoethanolamine transferase specific for the outer 3-deoxy-d-manno-octulosonic acid residue of Escherichia coli lipopolysaccharide. Identification of the eptB gene and Ca[sup]2+[/sup] hypersensitivity of an eptB deletion mutant. J Biol Chem 280:21202–21211 [CrossRef]
     [Google Scholar]
  26. Schweizer H. D. 1993; Small broad-host-range gentamycin resistance gene cassettes for site-specific insertion and deletion mutagenesis. Biotechniques 15:831–834
     [Google Scholar]
  27. Shi Y, Cromie M. J, Hsu F. F, Turk J, Groisman E. A. 2004; PhoP-regulated Salmonella resistance to the antimicrobial peptides magainin 2 and polymyxin B. Mol Microbiol 53:229–241 [CrossRef]
     [Google Scholar]
  28. Somerville J. E, Cassiano L, Darveau R. P Jr. 1999; Escherichia coli msbB gene as a virulence factor and a therapeutic target. Infect Immun 67:6583–6590
     [Google Scholar]
  29. Tamayo R, Ryan S. S, McCoy A. J, Gunn J. S. 2002; Identification and genetic characterization of PmrA-regulated genes and genes involved in polymyxin B resistance in Salmonella enterica serovar Typhimurium. Infect Immun 70:6770–6778 [CrossRef]
     [Google Scholar]
  30. Tamayo R, Choudhury B, Septer A, Merighi M, Carlson R, Gunn J. S. 2005; Identification of cptA , a PmrA-regulated locus required for phosphoethanolamine modification of the Salmonella enterica serovar typhimurium lipopolysaccharide core. J Bacteriol 187:3391–3399 [CrossRef]
     [Google Scholar]
  31. Tatsuno I, Horie M, Abe H. 7 other authors 2001; toxB gene on pO157 of enterohemorrhagic Escherichia coli O157 : H7 is required for full epithelial cell adherence phenotype. Infect Immun 69:6660–6669 [CrossRef]
     [Google Scholar]
  32. Trent M. S. 2004; Biosynthesis, transport, and modification of lipid A. Biochem Cell Biol 82:71–86 [CrossRef]
     [Google Scholar]
  33. Trent M. S, Raetz C. R. 2002; Cloning of EptA, the lipid A phosphoethanolamine transferase associated with polymyxin resistance. Abstract from the 7th Conference of the International Endotoxin Society, Washington, DC. J Endotoxin Res 8:158
     [Google Scholar]
  34. Tsubery H, Ofek I, Cohen S, Fridkin M. 2001; N-terminal modifications of Polymyxin B nonapeptide and their effect on antibacterial activity. Peptides 22:1675–1681 [CrossRef]
     [Google Scholar]
  35. Vaara M. 1992; Agents that increase the permeability of the outer membrane. Microbiol Rev 56:395–411
     [Google Scholar]
  36. Vaara M, Porro M. 1996; Group of peptides that act synergistically with hydrophobic antibiotics against gram-negative enteric bacteria. Antimicrob Agents Chemother 40:1801–1805
     [Google Scholar]
  37. Winfield M. D, Groisman E. A. 2004; Phenotypic differences between Salmonella and Escherichia coli resulting from the disparate regulation of homologous genes. Proc Natl Acad Sci U S A 101:17162–17167 [CrossRef]
     [Google Scholar]
  38. Wosten M. M, Kox L. F, Chamnongpol S, Soncini F. C, Groisman E. A. 2000; A signal transduction system that responds to extracellular iron. Cell 103:113–125 [CrossRef]
     [Google Scholar]
  39. Yoon J. W, Minnich S. A, Ahn J. S, Park Y. H, Paszczynski A, Hovde C. J. 2004; Thermoregulation of the Escherichia coli O157 : H7 pO157 ecf operon and lipid A myristoyl transferase activity involves intrinsically curved DNA. Mol Microbiol 51:419–435 [CrossRef]
     [Google Scholar]
  40. Yoon J. W, Lim J. Y, Park Y. H, Hovde C. J. 2005; Involvement of the Escherichia coli O157 : H7 (pO157) ecf operon and lipid A myristoyl transferase activity in bacterial survival in the bovine gastrointestinal tract and bacterial persistence in farm water troughs. Infect Immun 73:2367–2378 [CrossRef]
     [Google Scholar]
  41. Zhou Z, Lin S, Cotter R. J, Raetz C. R. H. 1999; Lipid A modifications characteristic of Salmonella typhimurium are induced by NH[sub]4[/sub]VO[sub]3[/sub] in Escherichia coli K12. J Biol Chem 274:18503–18514 [CrossRef]
     [Google Scholar]
  42. Zhou Z, Ribeiro A. A, Lin S, Cotter R. J, Miller S. I, Raetz C. R. H. 2001; Lipid A modifications in polymyxin-resistant Salmonella typhimurium : PMRA-dependent 4-amino-4-deoxy-l-arabinose, and phosphoethanolamine incorporation. J Biol Chem 276:43111–43121 [CrossRef]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.28692-0
Loading
Phosphoethanolamine substitution in the lipid A of Escherichia coli O157 : H7 and its association with PmrC
Microbiology 152, 657 (2006); https://doi.org/10.1099/mic.0.28692-0
/content/journal/micro/10.1099/mic.0.28692-0
/content/journal/micro/10.1099/mic.0.28692-0
Loading

Data & Media loading...

Most cited 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