Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1992 Jun;60(6):2166–2173. doi: 10.1128/iai.60.6.2166-2173.1992

Molecular characterization of an RTX toxin determinant from Actinobacillus suis.

L L Burrows 1, R Y Lo 1
PMCID: PMC257139  PMID: 1587585

Abstract

RTX cytolysins are a family of calcium-dependent, pore-forming, secreted toxins found in a variety of gram-negative bacteria. The prototypical member of this family is the alpha-hemolysin of Escherichia coli. The RTX genetic determinants from seven members of the family Pasteurellaceae, Pasteurella haemolytica, Actinobacillus actinomycetemcomitans, and A. pleuropneumoniae serotypes 1,5,7, and 9 were previously cloned and sequenced. Using the leukotoxin determinant from P. haemolytica serotype A1 as a probe, we detected the presence of RTX-type determinants in Actinobacillus suis, A. equuli, and A. lignieresii of the family Pasteurellaceae. All three species elaborate proteins of approximately 104 to 110 kDa that are recognized by polyclonal antisera against the 104-kDa hemolysin of A. pleuropneumoniae serotype 1. An RTX determinant of A. suis isolate 3714 was cloned and sequenced and was found to be almost identical to the RTX determinant of A. pleuropneumoniae serotypes 5 and 9. In addition, the determinant is not composed of four contiguous genes, as had been reported for most other RTX determinants; instead, the genes encoding the two proteins responsible for secretion of the toxin are at a locus distinct from that containing the toxin structural and activation genes.

Full text

PDF
2166

Images in this article

2168Image
on p.2168
2169Image
on p.2169

Selected References

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

  1. Anderson C., Potter A. A., Gerlach G. F. Isolation and molecular characterization of spontaneously occurring cytolysin-negative mutants of Actinobacillus pleuropneumoniae serotype 7. Infect Immun. 1991 Nov;59(11):4110–4116. doi: 10.1128/iai.59.11.4110-4116.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bendixen P. H., Shewen P. E., Rosendal S., Wilkie B. N. Toxicity of Haemophilus pleuropneumoniae for porcine lung macrophages, peripheral blood monocytes, and testicular cells. Infect Immun. 1981 Sep;33(3):673–676. doi: 10.1128/iai.33.3.673-676.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bisgaard M., Piechulla K., Ying Y. T., Frederiksen W., Mannheim W. Prevalence of organisms described as Actinobacillus suis or haemolytic Actinobacillus equuli in the oral cavity of horses. Comparative investigations of strains obtained and porcine strains of A. suis sensu stricto. Acta Pathol Microbiol Immunol Scand B. 1984 Dec;92(6):291–298. doi: 10.1111/j.1699-0463.1984.tb02836.x. [DOI] [PubMed] [Google Scholar]
  4. Carter P. L., Marshall R. B., Jolly R. D. A haemolytic variant of Actinobacillus equuli causing an acute septicaemia in a foal. N Z Vet J. 1971 Nov;19(11):264–265. doi: 10.1080/00480169.1971.33979. [DOI] [PubMed] [Google Scholar]
  5. Chang Y. F., Young R., Struck D. K. Cloning and characterization of a hemolysin gene from Actinobacillus (Haemophilus) pleuropneumoniae. DNA. 1989 Nov;8(9):635–647. doi: 10.1089/dna.1.1989.8.635. [DOI] [PubMed] [Google Scholar]
  6. Chang Y. F., Young R., Struck D. K. The Actinobacillus pleuropneumoniae hemolysin determinant: unlinked appCA and appBD loci flanked by pseudogenes. J Bacteriol. 1991 Aug;173(16):5151–5158. doi: 10.1128/jb.173.16.5151-5158.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Devenish J., Rosendal S., Bossé J. T. Humoral antibody response and protective immunity in swine following immunization with the 104-kilodalton hemolysin of Actinobacillus pleuropneumoniae. Infect Immun. 1990 Dec;58(12):3829–3832. doi: 10.1128/iai.58.12.3829-3832.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Devenish J., Rosendal S., Bossé J. T., Wilkie B. N., Johnson R. Prevalence of seroreactors to the 104-kilodalton hemolysin of Actinobacillus pleuropneumoniae in swine herds. J Clin Microbiol. 1990 Apr;28(4):789–791. doi: 10.1128/jcm.28.4.789-791.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Felmlee T., Pellett S., Welch R. A. Nucleotide sequence of an Escherichia coli chromosomal hemolysin. J Bacteriol. 1985 Jul;163(1):94–105. doi: 10.1128/jb.163.1.94-105.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Forestier C., Welch R. A. Nonreciprocal complementation of the hlyC and lktC genes of the Escherichia coli hemolysin and Pasteurella haemolytica leukotoxin determinants. Infect Immun. 1990 Mar;58(3):828–832. doi: 10.1128/iai.58.3.828-832.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frey J., Meier R., Gygi D., Nicolet J. Nucleotide sequence of the hemolysin I gene from Actinobacillus pleuropneumoniae. Infect Immun. 1991 Sep;59(9):3026–3032. doi: 10.1128/iai.59.9.3026-3032.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Glaser P., Sakamoto H., Bellalou J., Ullmann A., Danchin A. Secretion of cyclolysin, the calmodulin-sensitive adenylate cyclase-haemolysin bifunctional protein of Bordetella pertussis. EMBO J. 1988 Dec 1;7(12):3997–4004. doi: 10.1002/j.1460-2075.1988.tb03288.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Guthmiller J. M., Kolodrubetz D., Cagle M. P., Kraig E. Sequence of the lktB gene from Actinobacillus actinomycetemcomitans. Nucleic Acids Res. 1990 Sep 11;18(17):5291–5291. doi: 10.1093/nar/18.17.5291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Guthmiller J. M., Kraig E., Cagle M. P., Kolodrubetz D. Sequence of the lktD gene from Actinobacillus actinomycetemcomitans. Nucleic Acids Res. 1990 Sep 11;18(17):5292–5292. doi: 10.1093/nar/18.17.5292. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Inzana T. J., Todd J., Ma J. N., Veit H. Characterization of a non-hemolytic mutant of Actinobacillus pleuropneumoniae serotype 5: role of the 110 kilodalton hemolysin in virulence and immunoprotection. Microb Pathog. 1991 Apr;10(4):281–296. doi: 10.1016/0882-4010(91)90012-y. [DOI] [PubMed] [Google Scholar]
  16. Jensen A. E., Bertram T. A. Morphological and biochemical comparison of virulent and avirulent isolates of Haemophilus pleuropneumoniae serotype 5. Infect Immun. 1986 Feb;51(2):419–424. doi: 10.1128/iai.51.2.419-424.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kamp E. M., van Leengoed L. A. Serotype-related differences in production and type of heat-labile hemolysin and heat-labile cytotoxin of Actinobacillus (Haemophilus) pleuropneumoniae. J Clin Microbiol. 1989 Jun;27(6):1187–1191. doi: 10.1128/jcm.27.6.1187-1191.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kim B. H., Phillips J. E., Atherton J. G. Actinobacillus suis in the horse. Vet Rec. 1976 Mar 20;98(12):239–239. doi: 10.1136/vr.98.12.239. [DOI] [PubMed] [Google Scholar]
  19. Koronakis V., Cross M., Senior B., Koronakis E., Hughes C. The secreted hemolysins of Proteus mirabilis, Proteus vulgaris, and Morganella morganii are genetically related to each other and to the alpha-hemolysin of Escherichia coli. J Bacteriol. 1987 Apr;169(4):1509–1515. doi: 10.1128/jb.169.4.1509-1515.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kraig E., Dailey T., Kolodrubetz D. Nucleotide sequence of the leukotoxin gene from Actinobacillus actinomycetemcomitans: homology to the alpha-hemolysin/leukotoxin gene family. Infect Immun. 1990 Apr;58(4):920–929. doi: 10.1128/iai.58.4.920-929.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Larsen J. L. Isolation of haemolytic Actinobacilli from horses. Acta Pathol Microbiol Scand B Microbiol Immunol. 1974 Jun;82(3):453–454. doi: 10.1111/j.1699-0463.1974.tb02351.x. [DOI] [PubMed] [Google Scholar]
  22. Lo R. Y., Shewen P. E., Strathdee C. A., Greer C. N. Cloning and expression of the leukotoxin gene of Pasteurella haemolytica A1 in Escherichia coli K-12. Infect Immun. 1985 Dec;50(3):667–671. doi: 10.1128/iai.50.3.667-671.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lo R. Y., Strathdee C. A., Shewen P. E. Nucleotide sequence of the leukotoxin genes of Pasteurella haemolytica A1. Infect Immun. 1987 Sep;55(9):1987–1996. doi: 10.1128/iai.55.9.1987-1996.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MacInnes J. I., Rosendal S. Analysis of major antigens of Haemophilus (Actinobacillus) pleuropneumoniae and related organisms. Infect Immun. 1987 Jul;55(7):1626–1634. doi: 10.1128/iai.55.7.1626-1634.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mair N. S., Randall C. J., Thomas G. W., Harbourne J. F., McCrea C. T., Cowl K. P. Actinobacillus suis infection in pigs. A report of four outbreaks and two sporadic cases. J Comp Pathol. 1974 Jan;84(1):113–119. doi: 10.1016/0021-9975(74)90033-4. [DOI] [PubMed] [Google Scholar]
  26. Møller K., Kilian M. V factor-dependent members of the family Pasteurellaceae in the porcine upper respiratory tract. J Clin Microbiol. 1990 Dec;28(12):2711–2716. doi: 10.1128/jcm.28.12.2711-2716.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rosendal S., Devenish J., MacInnes J. I., Lumsden J. H., Watson S., Xun H. Evaluation of heat-sensitive, neutrophil-toxic, and hemolytic activity of Haemophilus (Actinobacillus) pleuropneumoniae. Am J Vet Res. 1988 Jul;49(7):1053–1058. [PubMed] [Google Scholar]
  28. Rosendal S., Mittal K. R. Serological cross-reactivity between a porcine Actinobacillus strain and Haemophilus pleuropneumoniae. Can J Comp Med. 1985 Apr;49(2):164–170. [PMC free article] [PubMed] [Google Scholar]
  29. Smits M. A., Briaire J., Jansen R., Smith H. E., Kamp E. M., Gielkens A. L. Cytolysins of Actinobacillus pleuropneumoniae serotype 9. Infect Immun. 1991 Dec;59(12):4497–4504. doi: 10.1128/iai.59.12.4497-4504.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Spitznagel J., Jr, Kraig E., Kolodrubetz D. Regulation of leukotoxin in leukotoxic and nonleukotoxic strains of Actinobacillus actinomycetemcomitans. Infect Immun. 1991 Apr;59(4):1394–1401. doi: 10.1128/iai.59.4.1394-1401.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Strathdee C. A., Lo R. Y. Cloning, nucleotide sequence, and characterization of genes encoding the secretion function of the Pasteurella haemolytica leukotoxin determinant. J Bacteriol. 1989 Feb;171(2):916–928. doi: 10.1128/jb.171.2.916-928.1989. [DOI] [PMC free article] [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]
  33. Welch R. A., Hull R., Falkow S. Molecular cloning and physical characterization of a chromosomal hemolysin from Escherichia coli. Infect Immun. 1983 Oct;42(1):178–186. doi: 10.1128/iai.42.1.178-186.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES