1887

Abstract

The genes encoding type IV O antigen glucosylation were characterized from both and . The putative O antigen modification genes from , , were PCR-amplified and introduced into serotype Y strain SFL124. Immunogold labelling and phage sensitivity indicated the presence of both serotype Y and serotype 4a O antigens on the cell surface of the resulting recombinant SFL124 strains, suggesting that only partial serotype conversion was conferred by the genes. The type IV O antigen modification genes were then isolated and characterized from serotype 4a strain NCTC 8296. A 38 kb chromosomal fragment conferred complete conversion to serotype 4a when introduced into SFL124. Sequence analysis of the fragment revealed the presence of three genes, . DNAs homologous to bacteriophage and were located upstream of , suggesting that this region of the NCTC 8296 genome may have originated from a bacteriophage; however, a serotype-converting phage could not be induced from this strain nor from other strains used in this study. Comparison of the GtrIV and GtrIV (o443) proteins revealed that they are 41% identical and 63% similar, which is the highest degree of similarity reported among the O antigen glucosyltransferases.

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2001-04-01
2019-12-09
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References

  1. Adhikari, P., Allison, G., Whittle, B. & Verma, N. K. ( 1999; ). Serotype 1a O-antigen modification: molecular characterization of the genes involved and their novel organization in the Shigella flexneri chromosome. J Bacteriol 181, 4711-4718.
    [Google Scholar]
  2. Allison, G. A. & Verma, N. K. ( 2000; ). Serotype-converting bacteriophages and O-antigen modification in Shigella flexneri. Trends Microbiol 8, 17-23.[CrossRef]
    [Google Scholar]
  3. Bastin, D. A., Lord, A. & Verma, N. K. ( 1997; ). Cloning and analysis of the glucosyl transferase gene encoding type I antigen in Shigella flexneri. FEMS Microbiol Lett 156, 133-139.[CrossRef]
    [Google Scholar]
  4. Blattner, F. R., Plunkett, G., III, Bloch, C. A. & 14 other authors ( 1997; ). The complete genome sequence of Escherichia coli K-12. Science 277, 1453–1462.[CrossRef]
    [Google Scholar]
  5. Breton, C. & Imberty, A. ( 1999; ). Structure/function studies of glycosyltransferases. Curr Opin Struct Biol 9, 563-571.[CrossRef]
    [Google Scholar]
  6. Carlin, N. I. A. & Lindberg, A. A. ( 1987; ). Monoclonal antibodies specific for Shigella flexneri lipopolysaccharides: clones binding to type IV, V, and VI antigens, group 3,4 antigen, and an epitope common to all Shigella flexneri and Shigella dysenteriae type 1 strains. Infect Immun 55, 1412-1420.
    [Google Scholar]
  7. Clark, C. A., Beltrame, J. & Manning, P. A. ( 1991; ). The oac gene encoding a lipopolysaccharide O-antigen acetylase maps adjacent to the integrase-encoding gene on the genome of Shigella flexneri bacteriophage Sf6. Gene 107, 43-52.[CrossRef]
    [Google Scholar]
  8. Guan, S. & Verma, N. K. ( 1998; ). Serotype conversion of a Shigella flexneri candidate vaccine strain via a novel site-specific chromosome-integration system. FEMS Microbiol Lett 166, 79-87.[CrossRef]
    [Google Scholar]
  9. Guan, S., Bastin, D. A. & Verma, N. K. ( 1999; ). Functional analysis of the O antigen glucosylation gene cluster of Shigella flexneri bacteriophage SfX. Microbiology 145, 1263-1273.[CrossRef]
    [Google Scholar]
  10. Hanahan, D. ( 1983; ). Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166, 557-580.[CrossRef]
    [Google Scholar]
  11. Huan, P. T., Taylor, R., Lindberg, A. A. & Verma, N. K. ( 1995; ). Immunogenicity of the Shigella flexneri serotype Y (SFL124) vaccine strain expressing cloned glucosyl transferase gene of converting bacteriophage SfX. Microbiol Immunol 39, 467-472.[CrossRef]
    [Google Scholar]
  12. Huan, P. T., Whittle, B. L., Bastin, D. A., Lindberg, A. A. & Verma, N. K. ( 1997a; ). Shigella flexneri type-specific antigen V: cloning, sequencing and characterization of the glucosyl transferase gene of temperate bacteriophage SfV. Gene 195, 207-216.[CrossRef]
    [Google Scholar]
  13. Huan, P. T., Bastin, D. A., Whittle, B. L., Lindberg, A. A. & Verma, N. K. ( 1997b; ). Molecular characterisation of the genes involved in O-antigen modification, attachment, integration and excision in Shigella flexneri bacteriophage SfV. Gene 195, 217-227.[CrossRef]
    [Google Scholar]
  14. Kotloff, K. L., Winickoff, J. P., Ivanoff, B., Clemens, J. D., Swerdlow, D. L., Sansonetti, P. J., Adak, G. K. & Levine, M. M. ( 1999; ). Global burden of Shigella infections: implications for vaccine development and implementation of control strategies. Bull WHO 77, 651-666.
    [Google Scholar]
  15. Lindberg, A. A., Karnell, A., Stocker, B. A., Katakura, S., Sweiha, H. & Rienholt, F. P. ( 1988; ). Development of an auxotrophic oral live Shigella flexneri vaccine. Vaccine 6, 146-150.[CrossRef]
    [Google Scholar]
  16. Mavris, M., Manning, P. A. & Morona, R. ( 1997; ). Mechanism of bacteriophage SfII-mediated serotype conversion in Shigella flexneri. Mol Microbiol 26, 939-950.[CrossRef]
    [Google Scholar]
  17. Morona, R., MacPherson, D. F., Van Den Bosch, L., Carlin, N. I. A. & Manning, P. A. ( 1995; ). Lipopolysaccharide with an altered O-antigen produced by Escherichia coli K-12 harbouring mutated, cloned Shigella flexneri rfb genes. Mol Microbiol 18, 209-223.[CrossRef]
    [Google Scholar]
  18. Petrovskaya, V. G. & Licheva, T. A. ( 1982; ). A provisional chromosome map of Shigella and the regions related to pathogenicity. Acta Microbiol Acad Sci Hung 29, 41-53.
    [Google Scholar]
  19. Phalipon, A., Kaufmann, M., Michetti, P., Cavaillon, J.-M., Huerre, M., Sansonetti, P. & Kraehenbuhl, J.-P. ( 1995; ). Monoclonal immunoglobulin A antibody directed against serotype-specific epitope of Shigella flexneri lipopolysaccharide protects against murine experimental shigellosis. J Exp Med 182, 769-778.[CrossRef]
    [Google Scholar]
  20. Rost, B., Casadio, R., Fariselli, P. & Sander, C. ( 1995; ). Prediction of helical transmembrane segments at 95% accuracy. Protein Sci 4, 521-533.
    [Google Scholar]
  21. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  22. Schuch, R. & Maurelli, A. T. ( 1997; ). Virulence plasmid instability in Shigella flexneri 2a is induced by virulence gene expression. Infect Immun 65, 3686-3692.
    [Google Scholar]
  23. Simmons, D. A. R. & Romanowska, E. ( 1987; ). Structure and biology of Shigella flexneri O antigens. J Med Microbiol 23, 289-302.[CrossRef]
    [Google Scholar]
  24. Stevenson, G., Neal, B., Liu, D., Hobbs, M., Packer, N. H., Batley, M., Redmond, J. W., Lindquist, L. & Reeves, P. ( 1994; ). Structure of the O antigen of Escherichia coli K-12 and the sequence of its rfb gene cluster. J Bacteriol 176, 4144-4156.
    [Google Scholar]
  25. Verma, N. K., Brandt, J. M., Verma, D. J. & Lindberg, A. A. ( 1991; ). Molecular characterisation of the O-acetyl transferase gene of converting bacteriophage SF6 that adds group antigen 6 to Shigella flexneri. Mol Microbiol 5, 71-75.[CrossRef]
    [Google Scholar]
  26. Verma, N. K., Verma, D. K., Huan, P. T. & Lindberg, A. A. ( 1993; ). Cloning and sequencing of the glucosyl transferase-encoding gene from converting bacteriophage X (SfX) of Shigella flexneri. Gene 129, 99-101.[CrossRef]
    [Google Scholar]
  27. Yanisch-Perron, C., Vieira, J. & Messing, J. ( 1985; ). Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33, 103-119.[CrossRef]
    [Google Scholar]
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