1887

Abstract

The roles of the three ORFs HP0208, HP0159 and HP1416 in the biosynthesis of 26695 LPS were investigated in this study. These ORFs represent a paralogous family of genes with homology to the serovar Typhimurium (hereafter referred to as ) gene, which encodes an α-1,2-glycosyltransferase required for core LPS biosynthesis. HP0208 contains multiple tandem repeats of the dimer 5′GA at its 5′ end and its expression is predicted to be subject to phase variation. The number of 5′GA repeats present in this ORF was found to be non-permissive for the expression of HP0208 in the majority of strains examined. To determine a role for this ORF in LPS biosynthesis a non-phase-variable, constitutively expressed variant of HP0208 was constructed and introduced into the genome of 26695. Analysis of the LPS profile of this strain by Tricine-SDS-PAGE and immunoblotting with anti-Lewis Y antigen (Le) mAbs confirmed a role for HP0208 in the biosynthesis of core LPS. A role for HP0159 and HP1416 in the biosynthesis of core LPS was also established. Although homologous to , HP0208, HP0159 and HP1416 failed to complement an mutant, suggesting that these ORFs encode functionally different enzymes.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.45842-0
2005-08-01
2019-11-18
Loading full text...

Full text loading...

/deliver/fulltext/jmm/54/8/JM540801.html?itemId=/content/journal/jmm/10.1099/jmm.0.45842-0&mimeType=html&fmt=ahah

References

  1. Alm R.A., Ling L.S., Moir D.T. and 20 other authors ( 1999;). Genomic-sequence comparison of two unrelated isolates of the human gastric pathogen Helicobacter pylori. Nature 397, 176–180.[CrossRef]
    [Google Scholar]
  2. Altman, E., Smirnova, N., Li, J., Aubry, A. & Logan, S. M. ( 2003;). Occurrence of a nontypable Helicobacter pylori strain lacking Lewis blood group O antigens and DD-heptoglycan: evidence for the role of the core 1,6-glucan chain in colonization. Glycobiology 13, 777–783.[CrossRef]
    [Google Scholar]
  3. Apicella, M. A., Shero, M., Dudas, K. C., Stack, R. R., LaScolea, L. J., Murphy, T. F. & Mylotte, J. M. ( 1984;). Fimbriation of Haemophilus influenzae species isolated from the respiratory tract of adults. J Infect Dis 150, 40–43.[CrossRef]
    [Google Scholar]
  4. Appelmelk, B. J., Martin, S. L., Monteiro, M. A. & 10 other authors ( 1999;). Phase variation in Helicobacter pylori lipopolysaccharide due to changes in the lengths of poly(C) tracts in 3-fucosyltransferase genes. Infect Immun 67, 5361–5366.
    [Google Scholar]
  5. Appelmelk, B. J., Martino, M. C., Veenhof, E. & 7 other authors ( 2000;). Phase variation in H type I and Lewis a epitopes of Helicobacter pylori lipopolysaccharide. Infect Immun 68, 5928–5932.[CrossRef]
    [Google Scholar]
  6. Aspinall, G. O., Monteiro, M. A., Pang, H., Walsh, E. & Moran, A. P. ( 1996;). Lipopolysaccharides of Helicobacter pylori type strain NCTC 11637 (ATCC 43504): Structures of the O-antigen and core oligosaccharide regions. Biochemistry 35, 2489–2497.[CrossRef]
    [Google Scholar]
  7. Blaser, M. J. ( 1998;). Helicobacter pylori and gastric diseases. Br Med J 316, 1507–1510.[CrossRef]
    [Google Scholar]
  8. Coutinho, P. M. & Henrissat, B. ( 1999;). Carbohydrate-active enzymes: an integrated database approach. In Recent Advances in Carbohydrate Bioengineering, pp. 3–12. Edited by H. J. Gilbert, G. Davies, B. Henrissat & B. Svensson. Cambridge: The Royal Society of Chemistry.
  9. Edwards, N. J., Monteiro, M. A., Faller, G., Walsh, E. J., Moran, A. P., Roberts, I. S. & High, N. J. ( 2000;). Lewis X structures in the O-antigen side chain promote adhesion of Helicobacter pylori to the gastric epithelium. Mol Microbiol 35, 1530–1539.
    [Google Scholar]
  10. Falk, P., Roth, K. A., Boren, T., Westblom, T. U., Gordon, J. I. & Normark, S. ( 1993;). An in vitro adherence assay reveals that Helicobacter pylori exhibits lineage-specific tropism in the human gastric epithelium. Proc Natl Acad Sci U S A 90, 2035–2039.[CrossRef]
    [Google Scholar]
  11. Ferrero, R. L., Cussac, V., Courcoux, P. & Labigne, A. ( 1992;). Construction of isogenic urease-negative mutants of Helicobacter pylori by allelic exchange. J Bacteriol 174, 4212–4217.
    [Google Scholar]
  12. Fomsgaard, A., Freudenberg, M. A. & Galanos, C. ( 1990;). Modification of the silver staining technique to detect lipopolysaccharide in polyacrylamide gels. J Clin Microbiol 28, 2627–2631.
    [Google Scholar]
  13. Ge, Z., Chan, N. W. C., Palcic, M. M. & Taylor, D. E. ( 1997;). Cloning and heterologous expression of an 1,3 fucosyltransferase gene from the gastric pathogen Helicobacter pylori. J Biol Chem 272, 21357–21363.[CrossRef]
    [Google Scholar]
  14. Gottesman, C. S., Halpern, E. & Trisler, P. ( 1981;). Role of sulA and sulB in filamentation by Lon mutants of Escherichia coli K-12. J Bacteriol 148, 265–273.
    [Google Scholar]
  15. Heinrichs, D. E., Montiero, M. A., Perry, M. B. and Whitfield, C. ( 1998;). The assembly system for the lipopolysaccharide R2 core-type of Escherichia coli K-12 and Salmonella enterica.Structure and funciton of WaaK and WaaL homologues. J Biol Chem 273, 8849–8859.[CrossRef]
    [Google Scholar]
  16. Kimura, A. & Hansen, E. J. ( 1986;). Antigenic and phenotypic variants of Haemophilus influenzae type b lipopolysaccharide and their relationship to virulence. Infect Immun 51, 69–79.
    [Google Scholar]
  17. Lee, A., O'Rourke, J., De Ungria, M. C., Robertson, B., Daskalopoulos, G. & Dixon, M. F. ( 1997;). A standardised mouse model of Helicobacter pylori infection: introducing the Sydney strain. Gastroenterology 112, 1386–1397.[CrossRef]
    [Google Scholar]
  18. MacLachlan, P. R., Kadam, S. K. & Sanderson, K. E. ( 1991;). Cloning, characterisation, and DNA sequence of the rfa region for lipopolysaccharide synthesis in Salmonella typhimurium LT2. J Bacteriol 173, 7151–7163.
    [Google Scholar]
  19. Marshall, B. J. & Warren, J. R. ( 1984;). Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet i, 1311–1315.
    [Google Scholar]
  20. Martin, S. L., Edbrooke, M. R., Hodgman, T. C., van den Eijnden, D. H. & Bird, M. I. ( 1997;). Lewis X biosynthesis in Helicobacter pylori.Molecular cloning of an alpha(1,3)-fucosyltransferase gene. J Biol Chem 272, 21349–21356.[CrossRef]
    [Google Scholar]
  21. Monteiro, M. A. ( 2001;). Helicobacter pylori: a wolf in sheep's clothing: the glycotype families of Helicobacter pylori lipopolysaccharides expressing histo-blood groups: structure, biosynthesis, and role in pathogenesis. Adv Carbohydr Chem Biochem 57, 99–158.
    [Google Scholar]
  22. Monteiro, M. A., Appelmelk, B. J., Rasko, D. A. & 10 other authors ( 2000;). Lipopolysaccharide structures of Helicobacter pylori genomic strains 26695 and J99, mouse model H.pylori Sydney strain, H. pylori P466 carrying sialyl Lewis X, and H. pylori UA915 expressing Lewis B classification of H. pylori lipopolysaccharides into glycotype families. Eur J Biochem 267, 305–320.[CrossRef]
    [Google Scholar]
  23. Patrick, C. C., Patrick, G. S., Kaplan, S. L., Barrish, J. & Mason, E. O. ( 1989;). Adherence kinetics of Haemophilus influenzae type b to eucaryotic cells. Pediatr Res 26, 500–503.[CrossRef]
    [Google Scholar]
  24. Roantree, R. J., Kuo, T. T. & MacPhee, D. G. ( 1977;). The effect of defined lipopolysaccharide core defects upon antibiotic resistances of Salmonella typhimurium. J Gen Microbiol 103, 223–234.[CrossRef]
    [Google Scholar]
  25. Salaun, L., Linz, B., Suerbaum, S. & Saunders, N. J. ( 2004;). The diversity within an expanded and redefined repertoire of phase-variable genes in Helicobacter pylori. Microbiology 150, 817–830.[CrossRef]
    [Google Scholar]
  26. Sambrook, J. & Russell, D. W. ( 2001;). Transformation of E. coli by electroporation. In Molecular Cloning: a Laboratory Manual, pp. 1–119. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  27. Schagger, H. & von Jagow, G. ( 1987;). Tricine-sodium dodecylsulphate-polyacrylamide gel electrophoresis for the separation of proteins in the range of 1 to 100 kDa. Anal Biochem 166, 368–379.[CrossRef]
    [Google Scholar]
  28. Shibayama, K., Ohsuka, S., Tanaka, T., Arakawa, Y. & Ohta, M. ( 1998;). Conserved structural regions involved in the catalytic mechanism of Escherichia coli K-12 WaaO (RfaI). J Bacteriol 180, 5313–5318.
    [Google Scholar]
  29. Tomb, J. F., White, O., Kerlavage, A. R. & 39 other authors ( 1997;). The complete genome sequence of the gastric pathogen Helicobacter pylori. Nature 388, 539–547.[CrossRef]
    [Google Scholar]
  30. van Ham, S. M., van Alphen, L., Mooi, F. R. & van Putten, J. P. ( 1993;). Phase variation of H.influenzae fimbriae: transcriptional control of two divergent genes through a variable combined promoter region. Cell 73, 1187–1196.[CrossRef]
    [Google Scholar]
  31. van Putten, J. P. ( 1993;). Phase variation of lipopolysaccharide directs the interconversion of invasive and immuno-resistant phenotypes of Neisseria gonorrhoeae. EMBO J 12, 4043–4051.
    [Google Scholar]
  32. Wang, Y. & Taylor, D. E. ( 1990;). Chloramphenicol resistance in Campylobacter coli: nucleotide sequence, expression and cloning vector construction. Gene 94, 23–28.[CrossRef]
    [Google Scholar]
  33. Wang, G., Boulton, P. G., Chan, N. W., Palcic, M. M. & Taylor, D. E. ( 1999;). Novel Helicobacter pylori alpha1,2-fucosyltransferase, a key enzyme in the synthesis of Lewis antigens. Microbiology 145, 3245–3253.
    [Google Scholar]
  34. Weiser, J. N. & Pan, N. ( 1998;). Adaptation of Haemophilus influenzae to acquired and innate humoral immunity based on phase variation of lipopolysaccharide. Mol Microbiol 30, 767–775.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.45842-0
Loading
/content/journal/jmm/10.1099/jmm.0.45842-0
Loading

Data & Media loading...

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