1887

Abstract

The Vi antigen is a capsular polysaccharide expressed by , the agent of human typhoid fever. Expression of this antigen is controlled by the and chromosomal loci. The locus is composed of 11 genes designated - (typhi Vi), - (Vi antigen export) and ORF11. We constructed Ty2 strains carrying non-polar mutations in ten genes located at the locus and examined the individual contribution of each gene to Vi phenotype. Phenotypes of the mutants and complementation experiments suggested that synthesis of Vi antigen monomer was catalysed by the TviB and TviC polypeptides. Subsequent polymerization of the polysaccharide might be catalysed by the TviE protein, but required functional TviD product. Proteins encoded by , and directed transport of the polymer to the bacterial cell surface. Anchoring of the Vi antigen at the bacterial cell surface was dependent of the VexE protein. The TviA protein was not essential for Vi polymer synthesis. However, disruption of the gene on Ty2 chromosome strongly decreased expression of Vi antigen. This defect was fully complemented by providing on a recombinant plasmid. By using transcriptional fusions, it was shown that the TviA product positively regulated co-transcription of the and genes from a promoter located upstream of . Moreover, we showed that a fusion was not expressed in a () mutant of . However, expression of the - fusion was restored in this mutant either by the gene of , or by the gene of when present in high copy number. This suggested that the and products could be involved in the same regulatory pathway modulating Vi antigen expression in . Together these results demonstrated that proteins encoded by the locus are not only involved in Vi polymer synthesis and translocation of the polysaccharide to the bacterial cell surface, but also in regulation of Vi antigen expression in .

Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-141-12-3039
1995-12-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/141/12/mic-141-12-3039.html?itemId=/content/journal/micro/10.1099/13500872-141-12-3039&mimeType=html&fmt=ahah

References

  1. Appleyard R.K. 1954; Segregation of new lysogenic types during growth of a double lysogenic strain derived from Escherichia coliK12.. Genetics 39:440–452
    [Google Scholar]
  2. Baker E.E., Whiteside R.E. 1965; Preparation and properties of a Vi antigen-degrading enzyme.. J Bacteriol 89:1217–1224
    [Google Scholar]
  3. Baker E.E., Whiteside R.E., Basch R., Derow M.A. 1959; The Vi antigen of the Enterobacteriaceae. II. Immunologic and biologic properties.. J Immunol 83:680–686
    [Google Scholar]
  4. Birnboim H.C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA.. Nucleic Acids Res 7:1513–1523
    [Google Scholar]
  5. Boulnois G.J., Roberts I.S. 1990; Genetics of capsular polysaccharide production in bacteria.. Curr Top Microbiol Immunol 150:1–18
    [Google Scholar]
  6. Boyer H.W., Roulland-Dussoix D.A. 1969; A complementation analysis of the restriction and modification of DNA in Escherichia coli.. J Mol Biol 41:459–472
    [Google Scholar]
  7. Casadaban M., Cohen S.N. 1980; Analysis of a gene control signal by DNA fusion and cloning in E. coli.. J Mol Biol 138:179–207
    [Google Scholar]
  8. Chang A.C.Y., Cohen S.N. 1978; Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from p15A cryptic miniplasmid.. J Bacteriol 134:1141–1155
    [Google Scholar]
  9. Churchward G., Belin D., Nagamine Y. 1984; A pSC101-derived plasmid which shows no sequence homology to other commonly used cloning vectors.. Gene 31:165–171
    [Google Scholar]
  10. Craigie J., Felix A. 1947; Typing of typhoid bacilli with Vi bacteriophage.. Lancet i824–827
    [Google Scholar]
  11. Daniels E.M., Schneerson R., Egan W.M., Szu S.C., Robbins J.B. 1989; Characterization of the Salmonella paratyphi C Vi polysaccharide.. Infect Immun 57:3159–3164
    [Google Scholar]
  12. Ditta G., Stanfield S., Corbin D., Helinski D.R. 1980; Broad host range DNA cloning system for Gram-negative bacteria: construction of a gene bank of Rhiɀobium meliloti.. Proc Natl Acad Sci USA 777347–7351
    [Google Scholar]
  13. Farinha M.A., Kropinski A.M. 1990; Construction of broad-host-range plasmid vectors for easy visible selection and analysis of promoters.. J Bacteriol 172:3496–3499
    [Google Scholar]
  14. Felix A., Pitt R.M. 1934; A new antigen of B. typhosus.. Lancet 227:186–191
    [Google Scholar]
  15. Felix A., Pitt R.M. 1936; The Vi antigens of various Salmonellatypes.. Br J Exp Pathol 17:81–86
    [Google Scholar]
  16. Frosch M., Müller A. 1993; Phospholipid substitution of capsular polysaccharides and mechanisms of capsule formation in Neisseria meningitidis.. Mol Microbiol 8:483–493
    [Google Scholar]
  17. Frosch M., Edwards U., Bousset K., Krauẞe B., Weisgerber C. 1991; Evidence for a common molecular origin of the capsule gene loci in Gram-negative bacteria expressing group II capsular polysaccharides.. Mol Microbiol 5:1251–1263
    [Google Scholar]
  18. Galan J.E., Ginocchio C., Costeas P. 1992; Molecular and functional characterization of the invasion gene invA: homology of InvA to members of a new protein family.. J Bacteriol 174:4338–4349
    [Google Scholar]
  19. Gottesman S., Stout V. 1991; Regulation of capsular poly-saccharide synthesis in Escherichia coli K-12.. Mol Microbiol 5:1599–1606
    [Google Scholar]
  20. Hashimoto Y., Ezaki T., Li N., Yamamoto H. 1991; Molecular cloning of the viaB region of Salmonella typhi.. FEMS Microbiol Lett 90:53–56
    [Google Scholar]
  21. Hashimoto Y., Li N., Yokoyama H., Ezaki T. 1993; Complete nucleotide sequence and molecular characterization of ViaB region encoding Vi antigen in Salmonella typhi.. J Bacteriol 175:4456–4465
    [Google Scholar]
  22. Heyns K., Kiessling G. 1967; Strukturaufklarung des Vi-antigens aus Citrobacter freundii (E. coli) 5396/38.. Carbohydr Res 3:340–352
    [Google Scholar]
  23. Higgins C.F. 1992; ABC transporters: from microorganisms to man.. Annu Rev Cell Biol 8:67–113
    [Google Scholar]
  24. Houng H.H., Noon K.F., Ou J.T., Baron L.S. 1992; Expression of Vi antigen in Escherichia coli K-12: characterization of ViaB from Citrobacter freundii and identity of ViaA with RcsB.. J Bacteriol 174:5910–5915
    [Google Scholar]
  25. Humphreys G.O., Weston A., Brown M.G.M., Saunders J.R. 1979; Plasmid transformation in Escherichia coli. . In Transformation pp. 254–279 Glover S.W., Buttler L.O. Edited by Oxford:: Cotswold Press.;
    [Google Scholar]
  26. Jann A., Jann K. 1990; Structure and biosynthesis of the capsular antigens of Escherichia coli.. Curr Top Microbiol Immunol 150:19–42
    [Google Scholar]
  27. Johnson E.M., Baron L.S. 1969; Genetic transfer of the Vi antigen from Salmonella typhosa to Escherichia coli.. J Bacteriol 99:355–359
    [Google Scholar]
  28. Johnson E.M., Krauskopf B., Baron L.S. 1965; Genetic mapping of Vi and somatic antigenic determinants in Salmonella.. J Bacteriol 90:302–308
    [Google Scholar]
  29. Johnson E.M., Krauskopf B., Baron L.S. 1966; Genetic analysis of the viaA-his chromosomal region in Salmonella.. J Bacteriol 92:1457–1463
    [Google Scholar]
  30. Kado C.I., Liu S.T. 1981; A rapid procedure for detection and isolation of large and small plasmids.. J Bacteriol 145:1365–1373
    [Google Scholar]
  31. Kolyva S., Waxin H., Popoff M.Y. 1992; The Vi antigen of Salmonella typhi: molecular analysis of the viaB locus.. J Gen Microbiol 138:297–304
    [Google Scholar]
  32. Kroll J.S., Loynds B., Brophy L.N., Moxon R. 1990; The hex locus in encapsulated Haemophilus influenzae: a chromosomal region involved in capsule polysaccharide export.. Mol Microbiol 4:1853–1862
    [Google Scholar]
  33. Le Minor L., Nicolle P. 1964; Sur deux souches de Salmonella dublin possedant l̓antigène Vi.. Ann Inst Pasteur 107:550–556
    [Google Scholar]
  34. Miller J.H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  35. Pickard D., Li J., Roberts M., Maskell D., Hone D., Levine M., Dougan G., Chatfield S. 1994; Characterization of defined ompR mutants of Salmonella typhi: ompR is involved in the regulation of Vi polysaccharide expression.. Infect Immun 62:3984–3993
    [Google Scholar]
  36. Sambrook J., Fritsch E.F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn.. Cold Spring Harbor, NY:: Cold Spring Harbor Laboratory.;
    [Google Scholar]
  37. Silver R.P., Aaronson W., Vann W.F. 1987; Translocation of capsular polysaccharides in pathogenic strains of Escherichia colirequires a 60-kilodalton periplasmic protein.. J Bacteriol 169:5489–5495
    [Google Scholar]
  38. Snellings N.J., Johnson E.M., Baron L.S. 1977; Genetic basis of Vi antigen expression in Salmonella paratyphi C.. J Bacteriol 131:57–62
    [Google Scholar]
  39. Stout V., Gottesman S. 1990; RcsB and RcsC: a two-component regulator of capsule synthesis in Escherichia coli.. J Bacteriol 172:659–669
    [Google Scholar]
  40. Vieira J., Messing J. 1982; The pUC plasmids, an Ml3 mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.. Gene 19:259–268
    [Google Scholar]
  41. Waxin H., Virlogeux I., Kolyva S., Popoff M.Y. 1993; Identification of six open reading frames in the Salmonella entericasubsp.enterica ser. Typhi viaB locus involved in Vi antigen production.. Res Microbiol 144:363–371
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-141-12-3039
Loading
/content/journal/micro/10.1099/13500872-141-12-3039
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