1887

Abstract

Almost all clinical enterohaemorrhagic (EHEC) O157: H7 isolates harbour a large virulence plasmid designated pO157. In this study, pO157 of EHEC O157:H7 reference strain EDL 933 was characterized at the molecular level. A restriction map was constructed by using seven restriction enzymes, with appropriate gel electrophoretic and hybridization methods. The molecular size of pO157 was determined to be 93.6 kb. By sequencing the DNA region extending in the 3′-direction of the previously described EHEC and genes, two further genes were discovered and analysed; these were termed EHEC and EHEC . The newly discovered genes together with the EHEC and genes constitute a typical RTX (Repeats in ToXin) determinant (EHEC operon) with the gene order . The map position of the operon was determined by hybridization experiments. Analysis of a DNA fragment carrying the downstream flanking region of the EHEC operon revealed an open reading frame which was highly homologous to of RepFIB, a basic replicon of IncF plasmids. It was located close to the EHEC operon.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-142-4-907
1996-04-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/4/mic-142-4-907.html?itemId=/content/journal/micro/10.1099/00221287-142-4-907&mimeType=html&fmt=ahah

References

  1. Benjamin M.M., Datta A.R. Acid tolerance of entero-hemorrhagic Escherichia coli. Appl Environ Microbiol 1995; 61:1669–1672
    [Google Scholar]
  2. Bhakdi S., Grimminger F., Suttorp N., Walmrath D., Seeger W. Proteinaceous bacterial toxins and pathogenesis of sepsis syndrome and septic shock: the unknown connection. Med Microbiol Immunol 1994; 183:119–144
    [Google Scholar]
  3. Coote J.G. Structural and functional relationships among the RTX toxin determinants of Gram-negative bacteria. FEMS Microbiol Rev 1992; 88:137–162
    [Google Scholar]
  4. Dytoc M., Soni R., Cockerill F., De Azavedo J., Louie M., Brunton J., Sherman P. Multiple determinants of verotoxin-producing Escherichia coli 0157 :H7 attachment and effacement. Infect Immun 1993; 61:3382–3391
    [Google Scholar]
  5. Fath M., J. & Kolter R. ABC transporters: bacterial exporters. Microbiol Rev 1993; 57:995–1017
    [Google Scholar]
  6. Fratamico P.M., Bhaduri S., Buchanan R.L. Studies on Escherichia coli serotype 0157:H7 strains containing a 60-MDa plasmid and on 60-MDa plasmid-cured derivatives. J Med Microbiol 1993; 39:371–381
    [Google Scholar]
  7. Gannon V.P., Teerling C., Masri S., A. & Gyles C.L. Molecular cloning and nucleotide sequence of another variant of the Escherichia coli Shiga-like toxin II family. J Gen Microbiol 1990; 136:1125–1135
    [Google Scholar]
  8. Gomez-Duarte O.G., Kaper J.B. A plasmid-encoded regulatory region activates chromosomal eaeA expression in enteropathogenic Escherichia coli. Infect Immun 1995; 63:1767–1776
    [Google Scholar]
  9. Griffin P.M., Tauxe R.V. The epidemiology of infections caused by Escherichia coli 0157:H7, other enterohemorrhagic E coli, and the associated hemolytic uremic syndrome. Epidemiol Rev 1991; 13:60–98
    [Google Scholar]
  10. Grinstedt J., Bennet P.M., Richmond M.H. A restriction enzyme map of the R-plasmid RP1. Plasmid 1977; 1:34–37
    [Google Scholar]
  11. Guthmiller J.M., Kolodrubetz D., Cagle M.P., Kraig E. Sequence of the IktB gene from Actinobacillus actino-mycetemcomitans. Nucleic Acids Res 1990a; 18:5291–1
    [Google Scholar]
  12. Guthmiller J.M., Kraig E., Cagle M.P., Kolodrubetz D. Sequence of the IktD gene from Actinobacillus actinomycetemcomitans. Nucleic Acids Res 1990b; 18:5292–1
    [Google Scholar]
  13. Guthmiller J.M., Kolodrubetz D., Kraig E. Mutational analysis of the putative leukotoxin transport genes in Actinobacillus actinomycetemcomitans. Microb Fat hog 1995; 18:307–321
    [Google Scholar]
  14. Hales B.A., Hart C.A., Batt R.M., Saunders J.R. The large plasmids found in enterohemorrhagic and enteropathogenic Escherichia coli constitute a related series of transfer-defective Inc F-IIA replicons. Plasmid 1992; Ll:183–193
    [Google Scholar]
  15. Heffron F. Tn3 and its relatives. In Mobile Genetic Elements 1983 Edited by Shapiro J.A. New York: Academic Press; pp 223–260
    [Google Scholar]
  16. Heffron F., McCarthy B.J., Ohtsubo H., Othsubo E. DNA sequence analysis of the transposon Tn3\ three genes and three sites involved in the transposition ofTn3. Cell 1979; 18:1153–1163
    [Google Scholar]
  17. Hess J., Wels W., Vogel M., Goebel W. Nucleotide sequence of a plasmid-encoded hemolysin determinant and its comparison with a corresponding chromosomal hemolysin sequence. FEMS Microbiol Eett 1986; 34:1–11
    [Google Scholar]
  18. Junkins A.D., Doyle M.P. Comparison of adherence properties of Escherichia coli of serotype 0157:H7 and a 60-megadalton plasmid-cured derivative. Curr Microbiol 1989; 19:21–27
    [Google Scholar]
  19. Karch H., Heesemann J., Laufs R., O'Brien A.D., Tacket C.O., Levine M.M. A plasmid of enterohemorrhagic Escherichia coli 0157:H7 is required for expression of a new fimbrial antigen and for adhesion to epithelial cells. Infect Immun 1987; 55:455–461
    [Google Scholar]
  20. Koronakis E., Hughes C., Milisav I., Koronakis V. Protein exporter function and in vitro ATPase activity are correlated in ABC-domain mutants of HlyB. Mol Microbiol 1995; 16:87–96
    [Google Scholar]
  21. Koronakis V., Hughes C., Koronakis E. ATPase activity and ATP/ADP-induced conformational change in the soluble domain of the bacterial protein translocator HlyB. Mol Microbiol 1993; 8:1163–1175
    [Google Scholar]
  22. Lally E.T., Golub E.E., Kieba I.R., Taichman N.S., Decker S., Berthold P., Gibson C.W., Demuth D.R., Rosenbloom J. Structure and function of the B and D genes of the Actinobacillus actinomycetemcomitans leukotoxin complex. Microb Pathog 1991; 11:111–121
    [Google Scholar]
  23. Louie M., De Azavedo J.C., Handelsman M.Y., Clark C.G., Ally B., Dytoc M., Sherman P., Brunton J. Expression and characterization of the eaeA gene product of Escherichia coli serotype 0157:H7. Infect Immun 1993; 61:4085–4092
    [Google Scholar]
  24. O'Brien A.D., Tesh V.L., Donohue Rolfe A., Jackson M.P., Olsnes S., Sandvig K., Lindberg A.A., Keusch G.T. Shiga toxin: biochemistry, genetics, mode of action, and role in pathogenesis. Curr Top Microbiol Immunol 1992; 180:65–94
    [Google Scholar]
  25. O'Brien A.D., Lively T.A., Chen M.E., Rothman S.W., Formal S.B. Escherichia coli 0157: H7 strains associated with haemorrhagic colitis in the United States produce a Shigella dysenteriae 1 (SHIGA) like cytotoxin. Lancet 1983; 1:702
    [Google Scholar]
  26. Oelschlaeger T.A., Barrett T.J., Kopecko D.J. Some structures and processes of human epithelial cells involved in uptake of enterohemorrhagic Escherichia coli 0157: H7 strains. Infect Immun 1994; 62:5142–5150
    [Google Scholar]
  27. Pridmore R.D. New and versatile cloning vectors with kanamycin-resistance marker. Gene 1987; 56:309–312
    [Google Scholar]
  28. Riley L.W., Remis R.S., Helgerson S.D., McGee H.B., Wells J.G., Davis B.R., Hebert R.J., Olcott E.S., Johnson L.M., Hargrett N.T., Blake P.A., Cohen M.L. Hemorrhagic colitis associated with a rare Escherichia coli serotype. N Engl J Med 1983; 308:681–685
    [Google Scholar]
  29. Sambrook J., Fritsch E.F., Maniatis T. Molecular Cloning: a Laboratory Manual, 2nd edn 1989 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Saul D., Spiers A.J., McAnulty J., Gibbs M.G., Bergquist P.L., Hill D.F. Nucleotide sequence and replication characteristics of RepFIB, a basic replicon of IncF plasmids. J Bacteriol 1989; 171:2697–2707
    [Google Scholar]
  31. Schmidt H., Plaschke B., Franke S., Rilssmann H., Schwarzkopf A., Heesemann J., Karch H. Differentiation in virulence patterns of Escherichia coli possessing eae genes. Med Microbiol Immunol 1993; 183:23–31
    [Google Scholar]
  32. Schmidt H., Karch H., Beutin L. The large-sized plasmids of enterohemorrhagic Escherichia coli 0157 strains encode hemolysins which are presumably members of the E coli a-hemolysin family. FEMS Microbiol Eett 1994; 117:189–196
    [Google Scholar]
  33. Schmidt H., Beutin L., Karch H. Molecular analysis of the plasmid-encoded hemolysin of Escherichia coli 0157: H7 strain EDL 933. Infect Immun 1995; 63:1055–1061
    [Google Scholar]
  34. Schuein R., Gentschev I., Mollenkopf H.J., Goebel W. A topological model for the haemolysin translocator protein HlyD. Mol Gen Genet 1992; 234:155–163
    [Google Scholar]
  35. Sherman P., Soni R., Petrie M., Karmali M. Surface properties of the Vero cytotoxin-producing Escherichia coli 0157: H7. Infect Immun 1987; 55:1824–1829
    [Google Scholar]
  36. Tarr P.I. Escherichia coli 0157: H7: clinical, diagnostic, and epidemiological aspects of human infection. Clin Infect Dis 1995; 20:1–10
    [Google Scholar]
  37. Tesh V.L., O'Brien A.D. The pathogenic mechanisms of Shiga toxin and the Shiga-like toxins. Mol Microbiol 1991; 5:1817–1822
    [Google Scholar]
  38. Toth I., Cohen M.L., Rumschlag H.S., Riley L.W., White I.E.H., Carr J.H., Bond W.W., Wachsmuth I.K. Influence of the 60-megadalton plasmid on adherence of Escherichia coli 0157: H7 and genetic derivatives. Infect Immun 1990; 58:1223–1231
    [Google Scholar]
  39. Tzipori S., Wachsmuth I.K., Chapman C., Birden R., Brittingham J., Jackson C., Hogg J. The pathogenesis of hemorrhagic colitis caused by Escherichia coli 0157: H7 in gnotobiotic piglets. J Infect Dis 1986; 154:712–716
    [Google Scholar]
  40. Tzipori S., Karch H., Wachsmuth K.I., Robins Browne R.M., O'Brien A.D., Lior H., Cohen M.L., Smithers J., Levine M.M. Role of a 60-megadalton plasmid and Shiga-like toxins in the pathogenesis of infection caused by enterohemorrhagic Escherichia coli 0157: H7 in gnotobiotic piglets. Infect Immun 1987; 55:3117–3125
    [Google Scholar]
  41. Tzipori S., Gunzer F., Donnenberg M.S., De Montigny L., Kaper J.B., Donohue-Rolfe A. The role of the eae A gene in diarrhea and neurological complications in a gnotobiotic piglet model of enterohemorrhagic Escherichia coli infection. Infect Immun 1995; 63:3621–3627
    [Google Scholar]
  42. Wadolkowski E.A., Burris J.A., O'Brien A.D. Mouse model for colonization and disease caused by enterohemorrhagic Escherichia coli 0157: H7. Infect Immun 1990; 58:2438–2445
    [Google Scholar]
  43. Wandersman C., Delepelaire P. TolC, an Escherichia coli outer membrane protein required for hemolysin secretion. Proc Natl Acad Sei USA 1990; 87:4776–4780
    [Google Scholar]
  44. Welch R.A. Pore-forming cytolysins of gram-negative bacteria. Mol Microbiol 1991; 5:521–528
    [Google Scholar]
  45. Welch R.A. Phylogenetic analyses of the RTX toxin family. In Virulence Mechanisms of Bacterial Pathogens 1995 Edited by Roth J.A. Washington, DC: American Society for Microbiology; pp 195–206
    [Google Scholar]
  46. Wells J.G., Davis B.R., Wachsmuth I.K., Riley L.W., Remis R.S., Sokolow R., Morris G.K. Laboratory investigation of hemorrhagic colitis outbreaks associated with a rare Escherichia coli serotype. J Clin Microbiol 1983; 18:512–520
    [Google Scholar]
  47. Yu J., Kaper J.B. Cloning and characterization of the eae gene of enterohaemorrhagic Escherichia coli 0157: H7. Mol Microbiol 1992; 6:411–417
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-142-4-907
Loading
/content/journal/micro/10.1099/00221287-142-4-907
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