1887

Abstract

Summary: A marine (designated sp. 60) that is related to was used as a host for a plasmid that encodes the non-toxic B subunit (EtxB) of heat-labile enterotoxin. Expression of EtxB in sp. 60 resulted in the efficient and selective secretion of the B subunit into the extracellular growth medium. This indicated that sp. 60, which does not normally produce cholera-like enterotoxins, nonetheless possesses a secretory machinery that permits these toxins to be translocated across its cytoplasmic and outer membranes. Expression of EtxB in a mutant of sp. 60 (MVT1192), which had previously been shown to be defective in the secretion of several extracellular proteins, resulted in approximately 95% of the B subunit remaining entrapped within the periplasm of the bacterial cell envelope. This implies that the mutation in MVT1192 defines a locus that determines a common step in the secretion of extracellular proteins, including oligomeric toxins.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-138-4-719
1992-04-01
2022-01-17
Loading full text...

Full text loading...

/deliver/fulltext/micro/138/4/mic-138-4-719.html?itemId=/content/journal/micro/10.1099/00221287-138-4-719&mimeType=html&fmt=ahah

References

  1. BAGDASARIAN M. M. , AMANN E. , LURZ R. ,, RUCKERT., B. & BAGDASARIAN M. . 1983; Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors. Gene 26:273–282
    [Google Scholar]
  2. DALLAS W. S. . 1983; Conformity between heat-labile toxin genes from human and porcine enterotoxigenic Escherichia coli. Infection and Immunity 40:1072–1077
    [Google Scholar]
  3. FINKELSTEIN R. A. , LOSPALLUTO J.J. . 1970; Production of highly purified cholergen and choleragenoid. Journal of Infectious Diseases 121:S63–S72
    [Google Scholar]
  4. FOCARETA T. , MANNING P. A. . 1987; Extracellular proteins of Vibrio cholerae: molecular cloning, nucleotide sequence and characterization of the deoxyribonuclease (DNase) together with its periplasmic location in Escherichia coli Κ-12. Gene 53:31–40
    [Google Scholar]
  5. FURSTE J. P. , PANSEGRAU W. , FRANK R. , BLOCKER H. , SCHOLZ P. , BAGDASARIAN M. , LANKA E. . 1986; Molecular cloning of the RP4 primase region in a multi-host-range tacP expression vector. Gene 48:119–131
    [Google Scholar]
  6. HARDY S. J. S. , HOLMGREN J. , JOHANSSON S. , SANCHEZ J. , HIRST T. R. . 1988; Coordinated assembly of multisubunit proteins: oligomerization of bacterial enterotoxins in vivo and in vitro. Proceedings of the National Academy of Sciences of the United States of America 85:7109–7113
    [Google Scholar]
  7. HIRST T. R. . 1991 Assembly and secretion of oligomeric toxins in Gram negative bacteria. A Source Book of Bacterial Protein Toxins75–100 J. E. Alouf , and J. H. Free . London: Academic Press;
    [Google Scholar]
  8. HIRST T. R. , HOLMGREN J. . 1987a; Transient entry of enterotoxin subunits into the periplasm occurs during their secretion from Vibrio cholerae. Journal of Bacteriology 169:1037–1045
    [Google Scholar]
  9. HIRST T. R. , HOLMGREN J. . 1987b; Conformation of protein secreted across bacterial outer membranes: a study of enterotoxin translocation from Vibrio cholerae. Proceedings of the National Academy of Sciences of the United States of America 84:7418–7422
    [Google Scholar]
  10. HIRST T. R. , WELCH R. A. . 1988; Mechanisms for secretion of extracellular proteins by Gram negative bacteria. Trends in Biochemical Sciences 13:265–269
    [Google Scholar]
  11. HIRST T. R. , LEECE R. . 1991; The phenomenon of toxin secretion in vibrios and aeromonads. Experientia 47:429–431
    [Google Scholar]
  12. HIRST T. R. , SANCHEZ J. , KAPER J. Β. , HARDY S. J. S. , HOLMGREN J. . 1984a; Mechanism of toxin secretion by Vibrio cholerae investigated in strains harbouring plasmids that encode heat-labile enterotoxins of Escherichia coli. Proceedings of the National Academy of Sciences of the United States of America 81:7752–7756
    [Google Scholar]
  13. HIRST T. R. , RANDALL L. L. , HARDY S. J. S. . 1984b; Cellular location of heat-labile enterotoxin in Escherichia coli. Journal of Bacteriology 157:637–642
    [Google Scholar]
  14. HOFSTRA Η. , WITHOLT Β. . 1984; Kinetics of synthesis, processing, and membrane transport of heat-labile enterotoxin, a periplasmic protein in Escherichia coli. Journal of Biological Chemistry 259:15182–15187
    [Google Scholar]
  15. HOFSTRA Η. , WITHOLT B. . 1985; Heat-labile enterotoxin of Escherichia coli: kinetics of association of subunits into periplasmic holotoxin. Journal of Biological Chemistry 260:16037–16044
    [Google Scholar]
  16. HOLMES R. K. , VASIL M. L. , FINKELSTEIN R. A. . 1975; Studies on toxigenesis of Vibrio cholerae. III. Characterization of nontoxinogenic mutants in vitro and in experimental animals. Journal of Clinical Investigation 55:551–560
    [Google Scholar]
  17. ICHIGE Α. , OISHI K. , MIZUSHIMA S. . 1988; Isolation and characterization of mutants of a marine Vibrio strain that are defective in the production of extracellular proteins. Journal of Bacteriology 170:3537–3542
    [Google Scholar]
  18. JANDA J. M. , POWERS C. , BRYANT R. G. , ABBOT S. L. . 1988; Current perspectives on the epidemiology and pathogenesis of clinically significant Vibrio spp. Clinical Microbiology Reviews 1:245–267
    [Google Scholar]
  19. LEONG J. , VINAL A. C. , DALLAS W. S. . 1985; Nucleotide sequence comparison between heat-labile toxin B-subunit cistrons from Escherichia coli of human and porcine origin. Infection and Immunity 48:73–77
    [Google Scholar]
  20. MEKALANOS J. J. , SWARTZ D. J. , PEARSON G. D. N. , HARFORD Ν. , GROYNE F. , DE WILDE M. . 1983; Cholera toxin genes: nucleotide sequence, deletion analysis and vaccine development. Nature, London 306:551–557
    [Google Scholar]
  21. MERCURIO A. , MANNING P. A. . 1985; Cellular location and export of the soluble haemolysin of Vibrio cholerae El Tor. Molecular and General Genetics 200:472–475
    [Google Scholar]
  22. MILLER J. H. . 1974 Experiments in Molecular Genetics. Cold Spring Harbor Laboratory, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  23. NEILL R. J. , IVINS B. E. , HOLMES R. K. . 1983a; Synthesis and secretion of the plasmid encoded heat-labile enterotoxin of Escherichia coli in Vibrio cholerae. Science 221:289–291
    [Google Scholar]
  24. NEILL R. J. , TWIDDY E. M. , HOLMES R. K. . 1983b; Synthesis of plasmid-coded heat-labile enterotoxin in wild-type and hypertoxinogenic strains of Escherichia coli and in other genera of Enterobacteriaceae. Infection and Immunity 41:1056–1061
    [Google Scholar]
  25. NISHIBUCHI M. , KAPER J. B. . 1985; Nucleotide sequence of the thermostable direct hemolysin gene from Vibrio cholerae. Journal of Bacteriology 162:558–564
    [Google Scholar]
  26. OISHI K. , YOKOSHIMA S. , TOMIYAMA T. , AIDA Κ. . 1979; Exohemagglutinins: new products of vibrios. Applied and Environmental Microbiology 38:169–172
    [Google Scholar]
  27. PALVA E. T. , HIRST T. R. , HARDY S. J. S. , HOLMGREN J. , RANDALL L. L. . 1981; Synthesis of a precursor of the Β subunit of heat-labile enterotoxin in Escherichia coli. Journal of Bacteriology 146:325–330
    [Google Scholar]
  28. SANDKVIST M. , HIRST T. R. , BAGDASARIAN M. . 1987; Alterations at the carboxyl terminus change assembly and secretion properties of the Β subunit of Escherichia coli heat-labile enterotoxin. Journal of Bacteriology 169:4570–4576
    [Google Scholar]
  29. SIMON R. , PRIEFER U. , PUHLER A. . 1983; A broad-host range mobilization system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria. Biotechnology 1:784–791
    [Google Scholar]
  30. SIXMA T. K. , PRONK S. E. , KALK Κ. H. , WARTNA E. S. , VAN ZANTAN Β. A. M. , WITHOLT B. , HOL W. G. J. . 1991; Crystal structure of a cholera toxin-related heat-labile enterotoxin from E. coli. Nature, London 351:371–377
    [Google Scholar]
  31. SVENNERHOLM A. M. , HOLMGREN J. . 1978; Identification of Escherichia coli heat-labile enterotoxin by means of a ganglioside immunosorbent assay (GM 1-ELISA) procedure. Current Microbiology 1:19–23
    [Google Scholar]
  32. VIMR E. R. , LAWRISUK L. , GALLEN J. , KAPER J. B. . 1988; Cloning and expression of the Vibrio cholerae neuraminidase gene nanH in Escherichia coli. Journal of Bacteriology 170:1495–1504
    [Google Scholar]
  33. WONG K. R. , BUCKLEY J. T. . 1989; Proton motive force involved in protein export from Aeromonas salmonicida. Science 246:654–656
    [Google Scholar]
  34. WONG K. R. , MCLEAN D. M. , BUCKLEY J. T. . 1990; Cloned aerolysin of Aeromonas hydrophila is exported by a wild-type marine Vibrio strain but remains periplasmic in pleiotropic export mutants. Journal of Bacteriology 172:372–376
    [Google Scholar]
  35. YOUNG D. B. , BROADBENT D. A. . 1982; Biochemical characterization of extracellular proteases from Vibrio cholerae. Infection and Immunity 37:875–883
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-138-4-719
Loading
/content/journal/micro/10.1099/00221287-138-4-719
Loading

Data & Media loading...

Most cited this month 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