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. 1989 Aug;57(8):2350–2357. doi: 10.1128/iai.57.8.2350-2357.1989

Detection of listeriolysin, the thiol-dependent hemolysin in Listeria monocytogenes, Listeria ivanovii, and Listeria seeligeri.

M Leimeister-Wächter 1, T Chakraborty 1
PMCID: PMC313454  PMID: 2744850

Abstract

The listeriolysin gene from a weakly hemolytic but virulent strain of Listeria monocytogenes serotype 1/2a was cloned in Escherichia coli K-12. Recombinants were identified on the basis of their cross-reactivities to hyperimmune antisera raised against streptolysin O and listeriolysin. Low levels of hemolytic activity were detected in crude lysates of strains harboring the listeriolysin gene. In DNA hybridization studies with five DNA probes that encoded the listeriolysin gene and surrounding sequences, highly homologous listeriolysin genes were found to be present in the species L. monocytogenes, Listeria ivanovii, and Listeria seeligeri. Immunoblotting performed with affinity-purified antibody to listeriolysin allowed the detection of this protein in supernatants of all three species. This study demonstrates for the first time that listeriolysin is produced by L. seeligeri and documents the genetic homology between the various listeriolysins produced by Listeria spp. Sequences unique to the species L. monocytogenes were found to be located downstream of the listeriolysin gene. Furthermore, the restriction fragment length polymorphisms detected with probes flanking the hlyA gene may be useful epidemiological markers in identifying and distinguishing virulent Listeria strains from each other.

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Selected References

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

  1. Berche P., Gaillard J. L., Geoffroy C., Alouf J. E. T cell recognition of listeriolysin O is induced during infection with Listeria monocytogenes. J Immunol. 1987 Dec 1;139(11):3813–3821. [PubMed] [Google Scholar]
  2. Buysse J. M., Stover C. K., Oaks E. V., Venkatesan M., Kopecko D. J. Molecular cloning of invasion plasmid antigen (ipa) genes from Shigella flexneri: analysis of ipa gene products and genetic mapping. J Bacteriol. 1987 Jun;169(6):2561–2569. doi: 10.1128/jb.169.6.2561-2569.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chakraborty T., Goebel W. Recent developments in the study of virulence in Listeria monocytogenes. Curr Top Microbiol Immunol. 1988;138:41–58. [PubMed] [Google Scholar]
  4. Dominguez Rodriguez L., Vazquez Boland J. A., Fernandez Garayzabal J. F., Echalecu Tranchant P., Gomez-Lucia E., Rodriguez Ferri E. F., Suarez Fernandez G. Microplate technique to determine hemolytic activity for routine typing of Listeria strains. J Clin Microbiol. 1986 Jul;24(1):99–103. doi: 10.1128/jcm.24.1.99-103.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  6. Flamm R. K., Hinrichs D. J., Thomashow M. F. Introduction of pAM beta 1 into Listeria monocytogenes by conjugation and homology between native L. monocytogenes plasmids. Infect Immun. 1984 Apr;44(1):157–161. doi: 10.1128/iai.44.1.157-161.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gaillard J. L., Berche P., Mounier J., Richard S., Sansonetti P. In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun. 1987 Nov;55(11):2822–2829. doi: 10.1128/iai.55.11.2822-2829.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gaillard J. L., Berche P., Sansonetti P. Transposon mutagenesis as a tool to study the role of hemolysin in the virulence of Listeria monocytogenes. Infect Immun. 1986 Apr;52(1):50–55. doi: 10.1128/iai.52.1.50-55.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Geoffroy C., Gaillard J. L., Alouf J. E., Berche P. Purification, characterization, and toxicity of the sulfhydryl-activated hemolysin listeriolysin O from Listeria monocytogenes. Infect Immun. 1987 Jul;55(7):1641–1646. doi: 10.1128/iai.55.7.1641-1646.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kampelmacher E. H., van Noorle Jansen L. M. Isolation of Listeria monocytogenes from faeces of clinically healthy humans and animals. Zentralbl Bakteriol Orig. 1969;211(3):353–359. [PubMed] [Google Scholar]
  11. Kathariou S., Metz P., Hof H., Goebel W. Tn916-induced mutations in the hemolysin determinant affecting virulence of Listeria monocytogenes. J Bacteriol. 1987 Mar;169(3):1291–1297. doi: 10.1128/jb.169.3.1291-1297.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kathariou S., Rocourt J., Hof H., Goebel W. Levels of Listeria monocytogenes hemolysin are not directly proportional to virulence in experimental infections of mice. Infect Immun. 1988 Feb;56(2):534–536. doi: 10.1128/iai.56.2.534-536.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kaufmann S. H. Acquired resistance to facultative intracellular bacteria: relationship between persistence, cross-reactivity at the T-cell level, and capacity to stimulate cellular immunity of different Listeria strains. Infect Immun. 1984 Jul;45(1):234–241. doi: 10.1128/iai.45.1.234-241.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kehoe M., Timmis K. N. Cloning and expression in Escherichia coli of the streptolysin O determinant from Streptococcus pyogenes: characterization of the cloned streptolysin O determinant and demonstration of the absence of substantial homology with determinants of other thiol-activated toxins. Infect Immun. 1984 Mar;43(3):804–810. doi: 10.1128/iai.43.3.804-810.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kreft J., Funke D., Haas A., Lottspeich F., Goebel W. Production, purification and characterization of hemolysins from Listeria ivanovii and Listeria monocytogenes Sv4b. FEMS Microbiol Lett. 1989 Jan 15;48(2):197–202. doi: 10.1111/j.1574-6968.1989.tb03298.x. [DOI] [PubMed] [Google Scholar]
  16. Kuhn M., Kathariou S., Goebel W. Hemolysin supports survival but not entry of the intracellular bacterium Listeria monocytogenes. Infect Immun. 1988 Jan;56(1):79–82. doi: 10.1128/iai.56.1.79-82.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mengaud J., Vicente M. F., Chenevert J., Pereira J. M., Geoffroy C., Gicquel-Sanzey B., Baquero F., Perez-Diaz J. C., Cossart P. Expression in Escherichia coli and sequence analysis of the listeriolysin O determinant of Listeria monocytogenes. Infect Immun. 1988 Apr;56(4):766–772. doi: 10.1128/iai.56.4.766-772.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Parrisius J., Bhakdi S., Roth M., Tranum-Jensen J., Goebel W., Seeliger H. P. Production of listeriolysin by beta-hemolytic strains of Listeria monocytogenes. Infect Immun. 1986 Jan;51(1):314–319. doi: 10.1128/iai.51.1.314-319.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Portnoy D. A., Jacks P. S., Hinrichs D. J. Role of hemolysin for the intracellular growth of Listeria monocytogenes. J Exp Med. 1988 Apr 1;167(4):1459–1471. doi: 10.1084/jem.167.4.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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