Skip to main content
PMC home page
Infection and Immunity logoLink to Infection and Immunity
. 1992 Jun;60(6):2182–2187. doi: 10.1128/iai.60.6.2182-2187.1992

Gene block encoding production of cytotoxic necrotizing factor 1 and hemolysin in Escherichia coli isolates from extraintestinal infections.

V Falbo 1, M Famiglietti 1, A Caprioli 1
PMCID: PMC257141  PMID: 1587586

Abstract

Cytotoxic necrotizing factors (CNFs) are Escherichia coli protein toxins causing cell multinucleation and enlargement in tissue cultures and necrosis in rabbit skin. In E. coli isolates causing urinary tract infections in humans, the production of CNF1 is closely associated with hemolysin production. In this study, we obtained data suggesting that this phenotypic association is due to the genetic linkage of the determinants of the two toxins on the chromosome of uropathogenic E. coli. The genes encoding hemolysin and CNF1 were shown to be closely linked in a 37-kb cloned DNA fragment from an E. coli urinary tract isolate of serotype O4:K12:H5 (E-B35). A DNA region encoding CNF1 production but not hemolysin production was further subcloned as a 12-kb SalI-EcoRI fragment and used as a CNF1-specific gene probe. DNA hybridization experiments indicated that the CNF1 and hemolysin determinants were closely linked on the chromosomes of isolate E-B35 and six additional extraintestinal isolates belonging to serogroups O2, O4, O6, O22, O75, and O85.

Full text

PDF
2184

Images in this article

2184Image
on p.2184
2185Image
on p.2185
2185Image
on p.2185
2186Image
on p.2186

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bisicchia R., Ciammarughi R., Caprioli A., Falbo V., Ruggeri F. M. Toxin production and haemagglutination in strains of Escherichia coli from diarrhoea in Brescia, Italy. J Hyg (Lond) 1985 Oct;95(2):353–361. doi: 10.1017/s002217240006277x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blanco J., Alonso M. P., González E. A., Blanco M., Garabal J. I. Virulence factors of bacteraemic Escherichia coli with particular reference to production of cytotoxic necrotising factor (CNF) by P-fimbriate strains. J Med Microbiol. 1990 Mar;31(3):175–183. doi: 10.1099/00222615-31-3-175. [DOI] [PubMed] [Google Scholar]
  4. Blanco J., González E. A., García S., Blanco M., Regueiro B., Bernárdez I. Production of toxins by Escherichia coli strains isolated from calves with diarrhoea in galicia (north-western Spain). Vet Microbiol. 1988 Dec;18(3-4):297–311. doi: 10.1016/0378-1135(88)90095-8. [DOI] [PubMed] [Google Scholar]
  5. Caprioli A., Donelli G., Falbo V., Possenti R., Roda L. G., Roscetti G., Ruggeri F. M. A cell division-active protein from E. coli. Biochem Biophys Res Commun. 1984 Jan 30;118(2):587–593. doi: 10.1016/0006-291x(84)91343-3. [DOI] [PubMed] [Google Scholar]
  6. Caprioli A., Falbo V., Roda L. G., Ruggeri F. M., Zona C. Partial purification and characterization of an escherichia coli toxic factor that induces morphological cell alterations. Infect Immun. 1983 Mar;39(3):1300–1306. doi: 10.1128/iai.39.3.1300-1306.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Caprioli A., Falbo V., Ruggeri F. M., Baldassarri L., Bisicchia R., Ippolito G., Romoli E., Donelli G. Cytotoxic necrotizing factor production by hemolytic strains of Escherichia coli causing extraintestinal infections. J Clin Microbiol. 1987 Jan;25(1):146–149. doi: 10.1128/jcm.25.1.146-149.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Caprioli A., Falbo V., Ruggeri F. M., Minelli F., Orskov I., Donelli G. Relationship between cytotoxic necrotizing factor production and serotype in hemolytic Escherichia coli. J Clin Microbiol. 1989 Apr;27(4):758–761. doi: 10.1128/jcm.27.4.758-761.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cavalieri S. J., Bohach G. A., Snyder I. S. Escherichia coli alpha-hemolysin: characteristics and probable role in pathogenicity. Microbiol Rev. 1984 Dec;48(4):326–343. doi: 10.1128/mr.48.4.326-343.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Rycke J., González E. A., Blanco J., Oswald E., Blanco M., Boivin R. Evidence for two types of cytotoxic necrotizing factor in human and animal clinical isolates of Escherichia coli. J Clin Microbiol. 1990 Apr;28(4):694–699. doi: 10.1128/jcm.28.4.694-699.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Rycke J., Guillot J. F., Boivin R. Cytotoxins in non-enterotoxigenic strains of Escherichia coli isolated from feces of diarrheic calves. Vet Microbiol. 1987 Oct;15(1-2):137–150. doi: 10.1016/0378-1135(87)90139-8. [DOI] [PubMed] [Google Scholar]
  12. De Rycke J., Phan-Thanh L., Bernard S. Immunochemical identification and biological characterization of cytotoxic necrotizing factor from Escherichia coli. J Clin Microbiol. 1989 May;27(5):983–988. doi: 10.1128/jcm.27.5.983-988.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Evans D. J., Jr, Evans D. G., Höhne C., Noble M. A., Haldane E. V., Lior H., Young L. S. Hemolysin and K antigens in relation to serotype and hemagglutination type of Escherichia coli isolated from extraintestinal infections. J Clin Microbiol. 1981 Jan;13(1):171–178. doi: 10.1128/jcm.13.1.171-178.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Felmlee T., Pellett S., Lee E. Y., Welch R. A. Escherichia coli hemolysin is released extracellularly without cleavage of a signal peptide. J Bacteriol. 1985 Jul;163(1):88–93. doi: 10.1128/jb.163.1.88-93.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ferragut C., Leclerc H. Etude comparative des methodes de détermination du Tm de l'ADN bactérien. Ann Microbiol (Paris) 1976 Feb-Mar;127A(2):223–235. [PubMed] [Google Scholar]
  16. Fiorentini C., Arancia G., Caprioli A., Falbo V., Ruggeri F. M., Donelli G. Cytoskeletal changes induced in HEp-2 cells by the cytotoxic necrotizing factor of Escherichia coli. Toxicon. 1988;26(11):1047–1056. doi: 10.1016/0041-0101(88)90203-6. [DOI] [PubMed] [Google Scholar]
  17. Hacker J., Bender L., Ott M., Wingender J., Lund B., Marre R., Goebel W. Deletions of chromosomal regions coding for fimbriae and hemolysins occur in vitro and in vivo in various extraintestinal Escherichia coli isolates. Microb Pathog. 1990 Mar;8(3):213–225. doi: 10.1016/0882-4010(90)90048-u. [DOI] [PubMed] [Google Scholar]
  18. Hacker J., Hughes C. Genetics of Escherichia coli hemolysin. Curr Top Microbiol Immunol. 1985;118:139–162. doi: 10.1007/978-3-642-70586-1_8. [DOI] [PubMed] [Google Scholar]
  19. High N. J., Hales B. A., Jann K., Boulnois G. J. A block of urovirulence genes encoding multiple fimbriae and hemolysin in Escherichia coli O4:K12:H-. Infect Immun. 1988 Feb;56(2):513–517. doi: 10.1128/iai.56.2.513-517.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  21. Holland R. E. Some infectious causes of diarrhea in young farm animals. Clin Microbiol Rev. 1990 Oct;3(4):345–375. doi: 10.1128/cmr.3.4.345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Low D., David V., Lark D., Schoolnik G., Falkow S. Gene clusters governing the production of hemolysin and mannose-resistant hemagglutination are closely linked in Escherichia coli serotype O4 and O6 isolates from urinary tract infections. Infect Immun. 1984 Jan;43(1):353–358. doi: 10.1128/iai.43.1.353-358.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mackman N., Nicaud J. M., Gray L., Holland I. B. Secretion of haemolysin by Escherichia coli. Curr Top Microbiol Immunol. 1986;125:159–181. doi: 10.1007/978-3-642-71251-7_10. [DOI] [PubMed] [Google Scholar]
  25. McLaren I., Wray C. Another animal Escherichia coli cytopathic factor. Vet Rec. 1986 Dec 6;119(23):576–577. [PubMed] [Google Scholar]
  26. Monti-Bragadin C., Samer L., Rottini G. D., Pani B. The compatibility of Hly factor, a transmissible element which controls alpha-haemolysin production in Escherichia coli. J Gen Microbiol. 1975 Feb;86(2):367–369. doi: 10.1099/00221287-86-2-367. [DOI] [PubMed] [Google Scholar]
  27. Noegel A., Rdest U., Goebel W. Determination of the functions of hemolytic plasmid pHly152 of Escherichia coli. J Bacteriol. 1981 Jan;145(1):233–247. doi: 10.1128/jb.145.1.233-247.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Oswald E., De Rycke J. A single protein of 110 kDa is associated with the multinucleating and necrotizing activity coded by the Vir plasmid of Escherichia coli. FEMS Microbiol Lett. 1990 Mar 15;56(3):279–284. [PubMed] [Google Scholar]
  29. Smith H. W. A search for transmissible pathogenic characters in invasive strains of Escherichia coli: the discovery of a plasmid-controlled toxin and a plasmid-controlled lethal character closely associated, or identical, with colicine V. J Gen Microbiol. 1974 Jul;83(0):95–111. doi: 10.1099/00221287-83-1-95. [DOI] [PubMed] [Google Scholar]
  30. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  31. Svanborg Edén C., Hagberg L., Hanson L. A., Hull S., Hull R., Jodal U., Leffler H., Lomberg H., Straube E. Bacterial adherence--a pathogenetic mechanism in urinary tract infections caused by Escherichia coli. Prog Allergy. 1983;33:175–188. [PubMed] [Google Scholar]
  32. Welch R. A., Dellinger E. P., Minshew B., Falkow S. Haemolysin contributes to virulence of extra-intestinal E. coli infections. Nature. 1981 Dec 17;294(5842):665–667. doi: 10.1038/294665a0. [DOI] [PubMed] [Google Scholar]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES