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. 1992 Feb;174(3):947–952. doi: 10.1128/jb.174.3.947-952.1992

The expression of virulence genes in Listeria monocytogenes is thermoregulated.

M Leimeister-Wächter 1, E Domann 1, T Chakraborty 1
PMCID: PMC206174  PMID: 1732227

Abstract

The expression of listeriolysin, a major virulence factor of the gram-positive facultative intracellular pathogen Listeria monocytogenes, is positively regulated by a transcriptional activator, the prfA gene product. We had previously shown that mutations within the prfA gene lead to loss of listeriolysin production. In this communication, the regulation of expression of listeriolysin by a specific environmental condition, namely, temperature, was studied in wild-type strains of Listeria monocytogenes. We found that expression of the hemolysis phenotype was thermoregulated. A lisA::lacZ fusion was constructed, and its expression in the wild-type strain was studied at various growth temperatures. The results showed that the fusion beta-galactosidase activity was expressed only when cultures were grown at temperatures above 30 degrees C. This activity could be either specifically repressed or induced, depending on growth temperature. No change in activity was detected in a strain harboring a control beta-galactosidase fusion at the various growth temperatures tested. Northern (RNA) blot analysis of lisA-specific RNA transcripts showed that thermoregulation is manifested at the level of transcription. We also found that the transcription of other PrfA-regulated virulence genes in L. monocytogenes was similarly affected by growth temperature. Hence, as in other facultative intracellular pathogens, Shigella and Yersinia spp., temperature is an important cue in the induction of expression of virulence genes in L. monocytogenes. Our studies revealed that a higher level of regulation is imposed on the PrfA-mediated activation of virulence genes in pathogenic L. monocytogenes.

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  1. Chakraborty T., Leimeister-Wächter M., Domann E., Hartl M., Goebel W., Nichterlein T., Notermans S. Coordinate regulation of virulence genes in Listeria monocytogenes requires the product of the prfA gene. J Bacteriol. 1992 Jan;174(2):568–574. doi: 10.1128/jb.174.2.568-574.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Domann E., Leimeister-Wächter M., Goebel W., Chakraborty T. Molecular cloning, sequencing, and identification of a metalloprotease gene from Listeria monocytogenes that is species specific and physically linked to the listeriolysin gene. Infect Immun. 1991 Jan;59(1):65–72. doi: 10.1128/iai.59.1.65-72.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dorman C. J., Ni Bhriain N., Higgins C. F. DNA supercoiling and environmental regulation of virulence gene expression in Shigella flexneri. Nature. 1990 Apr 19;344(6268):789–792. doi: 10.1038/344789a0. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Fleming D. W., Cochi S. L., MacDonald K. L., Brondum J., Hayes P. S., Plikaytis B. D., Holmes M. B., Audurier A., Broome C. V., Reingold A. L. Pasteurized milk as a vehicle of infection in an outbreak of listeriosis. N Engl J Med. 1985 Feb 14;312(7):404–407. doi: 10.1056/NEJM198502143120704. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Geoffroy C., Gaillard J. L., Alouf J. E., Berche P. Production of thiol-dependent haemolysins by Listeria monocytogenes and related species. J Gen Microbiol. 1989 Mar;135(3):481–487. doi: 10.1099/00221287-135-3-481. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Kok J., Leenhouts K. J., Haandrikman A. J., Ledeboer A. M., Venema G. Nucleotide sequence of the cell wall proteinase gene of Streptococcus cremoris Wg2. Appl Environ Microbiol. 1988 Jan;54(1):231–238. doi: 10.1128/aem.54.1.231-238.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Kyhse-Andersen J. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 1984 Dec;10(3-4):203–209. doi: 10.1016/0165-022x(84)90040-x. [DOI] [PubMed] [Google Scholar]
  12. Leimeister-Wächter M., Chakraborty T. Detection of listeriolysin, the thiol-dependent hemolysin in Listeria monocytogenes, Listeria ivanovii, and Listeria seeligeri. Infect Immun. 1989 Aug;57(8):2350–2357. doi: 10.1128/iai.57.8.2350-2357.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Leimeister-Wächter M., Domann E., Chakraborty T. Detection of a gene encoding a phosphatidylinositol-specific phospholipase C that is co-ordinately expressed with listeriolysin in Listeria monocytogenes. Mol Microbiol. 1991 Feb;5(2):361–366. doi: 10.1111/j.1365-2958.1991.tb02117.x. [DOI] [PubMed] [Google Scholar]
  14. Leimeister-Wächter M., Haffner C., Domann E., Goebel W., Chakraborty T. Identification of a gene that positively regulates expression of listeriolysin, the major virulence factor of listeria monocytogenes. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8336–8340. doi: 10.1073/pnas.87.21.8336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Linnan M. J., Mascola L., Lou X. D., Goulet V., May S., Salminen C., Hird D. W., Yonekura M. L., Hayes P., Weaver R. Epidemic listeriosis associated with Mexican-style cheese. N Engl J Med. 1988 Sep 29;319(13):823–828. doi: 10.1056/NEJM198809293191303. [DOI] [PubMed] [Google Scholar]
  16. MACKANESS G. B. Cellular resistance to infection. J Exp Med. 1962 Sep 1;116:381–406. doi: 10.1084/jem.116.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Maurelli A. T. Temperature regulation of virulence genes in pathogenic bacteria: a general strategy for human pathogens? Microb Pathog. 1989 Jul;7(1):1–10. doi: 10.1016/0882-4010(89)90106-x. [DOI] [PubMed] [Google Scholar]
  18. Mengaud J., Geoffroy C., Cossart P. Identification of a new operon involved in Listeria monocytogenes virulence: its first gene encodes a protein homologous to bacterial metalloproteases. Infect Immun. 1991 Mar;59(3):1043–1049. doi: 10.1128/iai.59.3.1043-1049.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mengaud J., Vicente M. F., Cossart P. Transcriptional mapping and nucleotide sequence of the Listeria monocytogenes hlyA region reveal structural features that may be involved in regulation. Infect Immun. 1989 Dec;57(12):3695–3701. doi: 10.1128/iai.57.12.3695-3701.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Miller J. F., Mekalanos J. J., Falkow S. Coordinate regulation and sensory transduction in the control of bacterial virulence. Science. 1989 Feb 17;243(4893):916–922. doi: 10.1126/science.2537530. [DOI] [PubMed] [Google Scholar]
  21. Miller V. L., Mekalanos J. J. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Recsei P., Kreiswirth B., O'Reilly M., Schlievert P., Gruss A., Novick R. P. Regulation of exoprotein gene expression in Staphylococcus aureus by agar. Mol Gen Genet. 1986 Jan;202(1):58–61. doi: 10.1007/BF00330517. [DOI] [PubMed] [Google Scholar]
  24. Roy C. R., Miller J. F., Falkow S. The bvgA gene of Bordetella pertussis encodes a transcriptional activator required for coordinate regulation of several virulence genes. J Bacteriol. 1989 Nov;171(11):6338–6344. doi: 10.1128/jb.171.11.6338-6344.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sakai T., Sasakawa C., Yoshikawa M. Expression of four virulence antigens of Shigella flexneri is positively regulated at the transcriptional level by the 30 kiloDalton virF protein. Mol Microbiol. 1988 Sep;2(5):589–597. doi: 10.1111/j.1365-2958.1988.tb00067.x. [DOI] [PubMed] [Google Scholar]
  26. Schlech W. F., 3rd, Lavigne P. M., Bortolussi R. A., Allen A. C., Haldane E. V., Wort A. J., Hightower A. W., Johnson S. E., King S. H., Nicholls E. S. Epidemic listeriosis--evidence for transmission by food. N Engl J Med. 1983 Jan 27;308(4):203–206. doi: 10.1056/NEJM198301273080407. [DOI] [PubMed] [Google Scholar]
  27. Simpson W. J., LaPenta D., Chen C., Cleary P. P. Coregulation of type 12 M protein and streptococcal C5a peptidase genes in group A streptococci: evidence for a virulence regulon controlled by the virR locus. J Bacteriol. 1990 Feb;172(2):696–700. doi: 10.1128/jb.172.2.696-700.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sokolovic Z., Fuchs A., Goebel W. Synthesis of species-specific stress proteins by virulent strains of Listeria monocytogenes. Infect Immun. 1990 Nov;58(11):3582–3587. doi: 10.1128/iai.58.11.3582-3587.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sokolovic Z., Goebel W. Synthesis of listeriolysin in Listeria monocytogenes under heat shock conditions. Infect Immun. 1989 Jan;57(1):295–298. doi: 10.1128/iai.57.1.295-298.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Straley S. C., Bowmer W. S. Virulence genes regulated at the transcriptional level by Ca2+ in Yersinia pestis include structural genes for outer membrane proteins. Infect Immun. 1986 Feb;51(2):445–454. doi: 10.1128/iai.51.2.445-454.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wuenscher M. D., Köhler S., Goebel W., Chakraborty T. Gene disruption by plasmid integration in Listeria monocytogenes: insertional inactivation of the listeriolysin determinant lisA. Mol Gen Genet. 1991 Aug;228(1-2):177–182. doi: 10.1007/BF00282463. [DOI] [PubMed] [Google Scholar]
  32. van Netten P., Perales I., van de Moosdijk A., Curtis G. D., Mossel D. A. Liquid and solid selective differential media for the detection and enumeration of L. monocytogenes and other Listeria spp. Int J Food Microbiol. 1989 Jul;8(4):299–316. doi: 10.1016/0168-1605(89)90001-9. [DOI] [PubMed] [Google Scholar]

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