1887

Abstract

In previous studies the authors cloned and characterized the DNA sequence of the regions at both ends of the O7-specific lipopolysaccharide (LPS) biosynthesis cluster of VW187 (O7:K1), and identified the biosynthetic genes for dTDP-rhamnose and GDP-mannose, as well as one of the candidate glycosyltransferases. In this work the complete DNA sequence of a 69 kb intervening region is presented. Seven new ORFs were identified. All the functions required for the synthesis and transfer of the O7 LPS were assigned on the basis of complementation experiments of transposon insertion mutants, and amino acid sequence homology to proteins involved in LPS synthesis of other bacteria. Of the seven ORFs, two encoded membrane proteins that were homologous to the O-antigen translocase (Wzx) and polymerase (Wxy), two were involved in the biosynthesis of dTDP--acetylviosamine, and the remaining three showed homologies to sugar transferases. The O antigen chain length regulator gene was also identified in the vicinity of the O7 polysaccharide cluster. O7-specific DNA primers were designed and tested for serotyping of O7 strains.

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1999-09-01
2019-09-17
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References

  1. Alexander, D. C. & Valvano, M. A. ( 1994; ). Role of the rfe gene in the biosynthesis of the Escherichia coli O7-specific lipopolysaccharide and other O-specific polysaccharides containing N-acetylglucosamine. J Bacteriol 176, 7079-7084.
    [Google Scholar]
  2. Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. ( 1997; ). Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res 25, 3389-3402.[CrossRef]
    [Google Scholar]
  3. Baev, N. & Kondorosi, A. ( 1992; ). Nucleotide sequence of the Rhizobium meliloti nodL gene located in locus n5 of the nod regulon. Plant Mol Biol 18, 843-846.[CrossRef]
    [Google Scholar]
  4. Batchelor, R. A., Alifano, P., Biffali, E., Hull, S. I. & Hull, R. A. ( 1992; ). Nucleotide sequences of the genes regulating O-polysaccharide antigen chain length (rol) from Escherichia coli and Salmonella typhimurium: protein homology and functional complementation. J Bacteriol 174, 5228-5236.
    [Google Scholar]
  5. Berlyn, M. K. ( 1998; ). Linkage map of Escherichia coli K-12, edition 10: the traditional map. Microbiol Mol Biol Rev 62, 814-984.
    [Google Scholar]
  6. Bugert, P. & Geider, K. ( 1995; ). Molecular analysis of the ams operon required for exopolysaccharide synthesis in Erwinia amylovora. Mol Microbiol 15, 917-933.[CrossRef]
    [Google Scholar]
  7. Cohen, S. N., Chang, A. C. & Hsu, L. ( 1972; ). Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci USA 69, 2110-2114.[CrossRef]
    [Google Scholar]
  8. Cserzo, M., Wallin, E., Simon, I., von Heijne, G. & Elofsson, A. ( 1997; ). Prediction of transmembrane alpha-helices in procaryotic membrane proteins: the Dense Alignment Surface method. Protein Eng 10, 673-676.[CrossRef]
    [Google Scholar]
  9. Daniels, C., Vindurampulle, C. & Morona, R. ( 1998; ). Overexpression and topology of the Shigella flexneri O-antigen polymerase (Rfc/Wzy). Mol Microbiol 28, 1211-1222.[CrossRef]
    [Google Scholar]
  10. Dicker, I. B. & Seetharam, S. ( 1992; ). What is known about the structure and function of the Escherichia coli protein FirA? Mol Microbiol 6, 817-823.[CrossRef]
    [Google Scholar]
  11. Dietzler, D. N. & Strominger, J. L. ( 1973; ). Characterization of the 4-acetamido-4,6-dideoxyhexoses from Escherichia coli strains. J Biol Chem 248, 104-109.
    [Google Scholar]
  12. Dykxhoorn, D. M., St Pierre, R. & Linn, T. ( 1996; ). A set of compatible tac promoter expression vectors. Gene 177, 133-136.[CrossRef]
    [Google Scholar]
  13. 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]
  14. Henikoff, S. ( 1984; ). Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28, 351-359.[CrossRef]
    [Google Scholar]
  15. Joiner, K. A. ( 1988; ). Complement evasion by bacteria and parasites. Annu Rev Microbiol 42, 201-230.[CrossRef]
    [Google Scholar]
  16. Lin, W. S., Cunneen, T. & Lee, C. Y. ( 1994; ). Sequence analysis and molecular characterization of genes required for the biosynthesis of type 1 capsular polysaccharide in Staphylococcus aureus. J Bacteriol 176, 7005-7016.
    [Google Scholar]
  17. Liu, D. & Reeves, P. R. ( 1994; ). Escherichia coli K12 regains its O antigen. Microbiology 140, 49-57.[CrossRef]
    [Google Scholar]
  18. Liu, D., Cole, R. A. & Reeves, P. R. ( 1996; ). An O-antigen processing function for Wzx (RfbX): a promising candidate for O-unit flippase. J Bacteriol 178, 2102-2107.
    [Google Scholar]
  19. Lukomski, S., Hull, R. A. & Hull, S. I. ( 1996; ). Identification of the O antigen polymerase (rfc) gene in Escherichia coli O4 by insertional mutagenesis using a nonpolar chloramphenicol resistance cassette. J Bacteriol 178, 240-247.
    [Google Scholar]
  20. L’vov, V. L., Shashkiv, A. S., Dimitriev, B. A., Kochtkov, N. K., Jann, B. & Jann, K. ( 1984; ). Structural studies of the O-specific side chain of the lipopolysaccharide from Escherichia coli O:7. Carbohydr Res 126, 249-259.[CrossRef]
    [Google Scholar]
  21. McLaughlin, R., Spinola, S. M. & Apicella, M. A. ( 1992; ). Generation of lipooligosaccharide mutants of Haemophilus influenzae type b. J Bacteriol 174, 6455-6459.
    [Google Scholar]
  22. Marolda, C. L. & Valvano, M. A. ( 1993; ). Identification, expression, and DNA sequence of the GDP-mannose biosynthesis genes encoded by the O7 rfb gene cluster of strain VW187 (Escherichia coli O7:K1). J Bacteriol 175, 148-158.
    [Google Scholar]
  23. Marolda, C. L. & Valvano, M. A. ( 1995; ). Genetic analysis of the dTDP-rhamnose biosynthesis region of the Escherichia coli VW187 (O7:K1) rfb gene cluster: identification of functional homologs of rfbB and rfbA in the rff cluster and correct location of the rffE gene. J Bacteriol 177, 5539-5546.
    [Google Scholar]
  24. Marolda, C. L. & Valvano, M. A. ( 1996; ). The GalF protein of Escherichia coli is not an UDP-glucose pyrophosphorylase but interacts with the GalU protein to possibly regulate cellular levels of UDP-glucose. Mol Microbiol 22, 827-840.[CrossRef]
    [Google Scholar]
  25. Marolda, C. L. & Valvano, M. A. ( 1998; ). The promoter region of the Escherichia coli O7-specific lipopolysaccharide gene cluster: structural and functional characterization of an upstream untranslated mRNA sequence. J Bacteriol 180, 3070-3079.
    [Google Scholar]
  26. Marolda, C. L., Welsh, J., Dafoe, L. & Valvano, M. A. ( 1990; ). Genetic analysis of the O7-polysaccharide biosynthesis region from the Escherichia coli O7:K1 strain VW187. J Bacteriol 172, 3590-3599.
    [Google Scholar]
  27. Matsuhashi, M. & Strominger, J. L. ( 1964; ). Thymidine diphosphate 4-acetamido-4,6-dideoxyhexoses. I. Enzymatic synthesis by strains of Escherichia coli. J Biol Chem 239, 2454-2463.
    [Google Scholar]
  28. Matsuhashi, M. & Strominger, J. L. ( 1966; ). Thymidine diphosphate 4-acetamido-4,6-dideoxyhexoses. III. Purification and properties of thymidine diphosphate 4-keto-6-deoxy-d-glucose transaminase from Escherichia coli strain B. J Biol Chem 241, 4738-4744.
    [Google Scholar]
  29. Morona, R., Mavris, M., Fallarino, A. & Manning, P. A. ( 1994; ). Characterization of the rfc region of Shigella flexneri. J Bacteriol 176, 733-747.
    [Google Scholar]
  30. Nikaido, H. & Vaara, M. ( 1985; ). Molecular basis of bacterial outer membrane permeability. Microbiol Rev 49, 1-32.
    [Google Scholar]
  31. Ochman, H. & Lawrence, J. G. (1996). Phylogenetics and amelioration of bacterial genomes. In Escherichia coli and Salmonella. Cellular and Molecular Biology, pp. 2627–2637. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  32. Paulsen, I. T., Beness, A. M. & Saier, M. H. ( 1997; ). Computer-based analyses of the protein constituents of transport systems catalysing export of complex carbohydrates in bacteria. Microbiology 143, 2685-2699.[CrossRef]
    [Google Scholar]
  33. Pluschke, G. & Achtman, M. ( 1984; ). Degree of antibody-independent activation of the classical complement pathway by K1 Escherichia coli differs with O antigen type and correlates with virulence of meningitis in newborns. Infect Immun 43, 684-692.
    [Google Scholar]
  34. Raetz, C. R. H. (1996). Bacterial lipopolysaccharides: a remarkable family of bioactive molecules. In Escherichia coli and Salmonella. Cellular and Molecular Biology, pp. 1035–1063. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  35. Reeves, P. R. ( 1992; ). Variation in O-antigens, niche-specific selection and bacterial populations. FEMS Microbiol Lett 79, 509-516.
    [Google Scholar]
  36. Reeves, P. R., Hobbs, M., Valvano, M. A. & 8 other authors ( 1996; ). Bacterial polysaccharide synthesis and gene nomenclature. Trends Microbiol 4, 495–503.[CrossRef]
    [Google Scholar]
  37. Rick, P. D. & Silver, R. P. (1996). Enterobacterial common antigen and capsular polysaccharides. In Escherichia coli and Salmonella. Cellular and Molecular Biology, pp. 104–122. Edited by F. C. Neidhardt and others. Washington, DC: American Society for Microbiology.
  38. Sanger, F., Nicklen, S. & Coulson, A. R. ( 1977; ). DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74, 5463-5467.[CrossRef]
    [Google Scholar]
  39. Selvaraj, G., Fong, Y. C. & Iyer, V. N. ( 1984; ). A portable DNA sequence carrying the cohesive site (cos) of bacteriophage lambda and the mob (mobilization) region of the broad-host-range plasmid RK2: a module for the construction of new cosmids. Gene 32, 235-241.[CrossRef]
    [Google Scholar]
  40. Smith, A. R. W., Rastall, R. A., Rees, N. H. & Hignett, R. C. ( 1990; ). Structure of the extracellular polysaccharide of Erwinia amylovora: a preliminary report. Acta Hortic 273, 211-219.
    [Google Scholar]
  41. 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 cluster. J Bacteriol 176, 4144-4156.
    [Google Scholar]
  42. Stevenson, G., Andrianopoulos, K., Hobbs, M. & Reeves, P. R. ( 1996; ). Organization of the Escherichia coli K-12 gene cluster responsible for production of the extracellular polysaccharide colanic acid. J Bacteriol 178, 4885-4893.
    [Google Scholar]
  43. Stroeher, U. H., Karageorgos, L. E., Brown, M. H., Morona, R. & Manning, P. A. ( 1995; ). A putative pathway for perosamine biosynthesis is the first function encoded within the rfb region of Vibrio cholerae O1. Gene 166, 33-42.[CrossRef]
    [Google Scholar]
  44. Tennigkeit, J. & Matzura, H. ( 1991; ). Nucleotide sequence analysis of a chloramphenicol-resistance determinant from Agrobacterium tumefaciens and identification of its gene product. Gene 98, 113-116.[CrossRef]
    [Google Scholar]
  45. Thompson, J. D., Higgins, D. G. & Gibson, T. J. ( 1994; ). clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position specific gap penalties and weight matrix choice. Nucleic Acids Res 22, 4673-4680.[CrossRef]
    [Google Scholar]
  46. Thorson, J. S., Lo, S. F. & Liu, H.-W. ( 1993; ). Biosynthesis of 3,6-dideoxyhexoses: new mechanistic reflections upon 2,6-dideoxy, 4,6-dideoxy, and amino sugar construction. J Am Chem Soc 115, 6993-6994.[CrossRef]
    [Google Scholar]
  47. Vaara, M. ( 1982; ). Eight bacterial proteins, including UDP-N-acetylglucosamine acyltransferase (LpxA) and three other transferases of Escherichia coli, consist of a six-residue periodicity theme. FEMS Microbiol Lett 76, 249-254.
    [Google Scholar]
  48. Valvano, M. A. ( 1992; ). Pathogenicity and molecular genetics of O-specific side-chain lipopolysaccharides of Escherichia coli. Can J Microbiol 38, 711-719.[CrossRef]
    [Google Scholar]
  49. Valvano, M. A. & Crosa, J. H. ( 1984; ). Aerobactin iron transport genes commonly encoded by certain ColV plasmids occur in the chromosome of a human invasive strain of Escherichia coli K1. Infect Immun 46, 159-167.
    [Google Scholar]
  50. Valvano, M. A. & Crosa, J. H. ( 1989; ). Molecular cloning and expression in Escherichia coli K-12 of chromosomal genes determining the O7 lipopolysaccharide antigen of a human invasive strain of E. coli O7:K1. Infect Immun 57, 937-943.
    [Google Scholar]
  51. Valvano, M. A. & Marolda, C. L. ( 1991; ). Relatedness of O-specific lipopolysaccharide side chain genes from strains of Shigella boydii type 12 belonging to two clonal groups and from Escherichia coli O7:K1. Infect Immun 59, 3917-3923.
    [Google Scholar]
  52. Wang, L. & Reeves, P. R. ( 1998; ). Organization of the Escherichia coli O157 O antigen cluster and identification of its specific genes. Infect Immun 66, 3545-3551.
    [Google Scholar]
  53. Whitfield, C. ( 1995; ). Biosynthesis of lipopolysaccharide O antigens. Trends Microbiol 3, 178-185.[CrossRef]
    [Google Scholar]
  54. Whitfield, C. & Valvano, M. A. ( 1993; ). Biosynthesis and expression of cell-surface polysaccharides in gram-negative bacteria. Adv Microb Physiol 35, 135-246.
    [Google Scholar]
  55. Yao, Z. & Valvano, M. A. ( 1994; ). Genetic analysis of the O-specific lipopolysaccharide biosynthesis region (rfb) of Escherichia coli K-12 W3110: identification of genes that confer group 6 specificity to Shigella flexneri serotypes Y and 4a. J Bacteriol 176, 4133-4143.
    [Google Scholar]
  56. Yao, Z., Liu, H. & Valvano, M. A. ( 1992; ). Acetylation of O-specific lipopolysaccharides from Shigella flexneri 3a and 2a occurs in Escherichia coli K-12 carrying cloned S. flexneri 3a and 2a rfb genes. J Bacteriol 174, 7500-7508.
    [Google Scholar]
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