Skip to main content
NCBI home page
Infection and Immunity logoLink to Infection and Immunity
. 1991 Dec;59(12):4497–4504. doi: 10.1128/iai.59.12.4497-4504.1991

Cytolysins of Actinobacillus pleuropneumoniae serotype 9.

M A Smits 1, J Briaire 1, R Jansen 1, H E Smith 1, E M Kamp 1, A L Gielkens 1
PMCID: PMC259069  PMID: 1937809

Abstract

Cytolysin I (ClyI) and cytolysin II (ClyII), which are present in the culture supernatant of Actinobacillus pleuropneumoniae serotype 9, are thought to play an important role in the pathogenesis of pig pleuropneumonia. The purpose of this study was to clone and characterize the genetic determinants of these cytolysins. Cloning was accomplished by the screening of DNA libraries for the presence of cytolytic activity and for the presence of DNA sequences homologous to leukotoxin DNA of Pasteurella haemolytica. Both genetic determinants were found to be members of the RTX cytotoxin family. The ClyII determinant was characterized in more detail. It appeared that ClyII more closely resembled the leukotoxin of P. haemolytica than the alpha-hemolysin of Escherichia coli. The ClyII amino acid sequence was identical to a hemolysin gene sequence of A. pleuropneumoniae serotype 5; this finding indicates that the latter gene also codes for ClyII and not for ClyI, as has previously been suggested. The genetic organization of the ClyII determinant differed from the genetic organization of other RTX determinants. Genes responsible for secretion of ClyII were not contiguous with the toxin gene. Instead, secretion genes were present elsewhere in the genome. These secretion genes, however, belong to the ClyI operon. This indicates that the secretion genes of the ClyI operon are responsible for secretion of ClyI and ClyII.

Full text

PDF
4497

Images in this article

4498Image
on p.4498
4502Image
on p.4502
4502Image
on p.4502
4503Image
on p.4503

Selected References

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

  1. Bertram T. A. Actinobacillus pleuropneumoniae: molecular aspects of virulence and pulmonary injury. Can J Vet Res. 1990 Apr;54 (Suppl):S53–S56. [PubMed] [Google Scholar]
  2. 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]
  3. Devenish J., Rosendal S. Identification of the heat-labile hemolysin of Actinobacillus pleuropneumoniae serotype 1. Can J Vet Res. 1989 Apr;53(2):251–254. [PMC free article] [PubMed] [Google Scholar]
  4. Devenish J., Rosendal S., Johnson R., Hubler S. Immunoserological comparison of 104-kilodalton proteins associated with hemolysis and cytolysis in Actinobacillus pleuropneumoniae, Actinobacillus suis, Pasteurella haemolytica, and Escherichia coli. Infect Immun. 1989 Oct;57(10):3210–3213. doi: 10.1128/iai.57.10.3210-3213.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Frey J., Nicolet J. Hemolysin patterns of Actinobacillus pleuropneumoniae. J Clin Microbiol. 1990 Feb;28(2):232–236. doi: 10.1128/jcm.28.2.232-236.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Frey J., Nicolet J. Regulation of hemolysin expression in Actinobacillus pleuropneumoniae serotype 1 by Ca2+. Infect Immun. 1988 Oct;56(10):2570–2575. doi: 10.1128/iai.56.10.2570-2575.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Frey J., Perrin J., Nicolet J. Cloning and expression of a cohemolysin, the CAMP factor of Actinobacillus pleuropneumoniae. Infect Immun. 1989 Jul;57(7):2050–2056. doi: 10.1128/iai.57.7.2050-2056.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gygi D., Nicolet J., Frey J., Cross M., Koronakis V., Hughes C. Isolation of the Actinobacillus pleuropneumoniae haemolysin gene and the activation and secretion of the prohaemolysin by the HlyC, HlyB and HlyD proteins of Escherichia coli. Mol Microbiol. 1990 Jan;4(1):123–128. doi: 10.1111/j.1365-2958.1990.tb02021.x. [DOI] [PubMed] [Google Scholar]
  10. Kamp E. M., Popma J. K., Anakotta J., Smits M. A. Identification of hemolytic and cytotoxic proteins of Actinobacillus pleuropneumoniae by use of monoclonal antibodies. Infect Immun. 1991 Sep;59(9):3079–3085. doi: 10.1128/iai.59.9.3079-3085.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Konings R. N., Verhoeven E. J., Peeters B. P. pKUN, vectors for the separate production of both DNA strands of recombinant plasmids. Methods Enzymol. 1987;153:12–34. doi: 10.1016/0076-6879(87)53045-2. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. MacInnes J. I., Kim J. E., Lian C. J., Soltes G. A. Actinobacillus pleuropneumoniae hlyX gene homology with the fnr gene of Escherichia coli. J Bacteriol. 1990 Aug;172(8):4587–4592. doi: 10.1128/jb.172.8.4587-4592.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Rycroft A. N., Williams D., Cullen J. M., Macdonald J. The cytotoxin of Actinobacillus pleuropneumoniae (pleurotoxin) is distinct from the haemolysin and is associated with a 120 kDa polypeptide. J Gen Microbiol. 1991 Mar;137(3):561–568. doi: 10.1099/00221287-137-3-561. [DOI] [PubMed] [Google Scholar]
  19. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sebunya T. N., Saunders J. R. Haemophilus pleuropneumoniae infection in swine: a review. J Am Vet Med Assoc. 1983 Jun 15;182(12):1331–1337. [PubMed] [Google Scholar]
  21. 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]
  22. Strathdee C. A., Lo R. Y. Extensive homology between the leukotoxin of Pasteurella haemolytica A1 and the alpha-hemolysin of Escherichia coli. Infect Immun. 1987 Dec;55(12):3233–3236. doi: 10.1128/iai.55.12.3233-3236.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Strathdee C. A., Lo R. Y. Regulation of expression of the Pasteurella haemolytica leukotoxin determinant. J Bacteriol. 1989 Nov;171(11):5955–5962. doi: 10.1128/jb.171.11.5955-5962.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES