1887

Abstract

Insertional mutagenesis was performed with Tn in the genetic background of an deletion mutant to identify new adhesion determinants in . Four insertion mutants defective in adhesion to eukaryotic cells were identified. Insertion sites were cloned by inverse-PCR and sequenced. The genetic organization of insertion regions was further analysed by screening and sequencing DNA fragments from a dIII library and by searching databases. Three adhesion-defective mutants each had one copy of Tn inserted into their chromosome. The insertion sites were different in the three mutants: (i) upstream from two ORFs in tandem, similar to and of , respectively; (ii) within an ORF encoding a putative 126 amino-acid-polypeptide with no significant similarity to any known protein; (iii) within an ORF similar to a ORF with no known function, just upstream from an operon similar to an ABC (ATP-binding cassette) transporter operon from . The excisants obtained from these mutants using the excision reporter plasmid pTCR9 recovered full adhesion capacity. A fourth mutant was the most severely defective in adhesion. It had five Tn insertions, one of which was upstream from and , and another of which was upstream from , a gene encoding a surface-exposed autolysin with a C terminus similar to that of InlB. Ami was clearly involved because an null mutant constructed in an EGDΔ background was adhesion-defective. Thus new regions involved in the adhesion of to eukaryotic cells were identified. Further study is required to define more accurately the roles of these regions in the adhesion process itself.

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2000-03-01
2020-01-21
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References

  1. Alvarez-Dominguez C., Carrasco-Marin E., Leyva-Cobian F.. 1993; Role of complement component C1q in phagocytosis of Listeria monocytogenes by murine macrophage-like cell lines. Infect Immun61:3664–3672
    [Google Scholar]
  2. Alvarez-Dominguez C., Vazquez-Boland J. A., Carrasco-Marin E., Lopez-Mato P., Leyva-Cobian F.. 1997; Host cell heparan sulfate proteoglycans mediate attachment and entry of Listeria monocytogenes, and the listerial surface protein ActA is involved in heparan sulfate receptor recognition. Infect Immun65:78–88
    [Google Scholar]
  3. Audurier A., Chatelain R., Chalons F., Piechaud M.. 1979; Lysotypie de 823 souches de Listeria monocytogenes isolées en France de 1958 à 1978. Ann Microbiol130B:179–189
    [Google Scholar]
  4. Birnboim H. C., Doly J.. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res7:1513–1523[CrossRef]
    [Google Scholar]
  5. Bradford M. M.. 1976; A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem72:248–254[CrossRef]
    [Google Scholar]
  6. Braun L., Dramsi S., Dehoux P., Bierne H., Lindahl G., Cossart P.. 1997; InlB: an invasion protein of Listeria monocytogenes with a novel type of surface association. Mol Microbiol25:285–294[CrossRef]
    [Google Scholar]
  7. Celli J., Poyart C., Trieu-Cuot P.. 1997; Use of an excision reporter plasmid to study the intracellular mobility of the conjugative transposon Tn916 in Gram-positive bacteria. Microbiology143:1253–1261[CrossRef]
    [Google Scholar]
  8. Cowart R. E., Lashmet J., McIntosh M. E., Adams T. J.. 1990; Adherence of a virulent strain of Listeria monocytogenes to the surface of a hepatocarcinoma cell line via lectin–substrate interaction. Arch Microbiol153:282–286[CrossRef]
    [Google Scholar]
  9. Dramsi S., Dehoux P., Cossart P.. 1993a; Common features of Gram-positive bacterial proteins involved in cell recognition. Mol Microbiol9:1119–1122[CrossRef]
    [Google Scholar]
  10. Dramsi S., Kocks C., Forestier C., Cossart P.. 1993b; Internalin-mediated invasion of epithelial cells by Listeria monocytogenes is regulated by bacterial growth state, temperature and the pleiotropic activator prfA. Mol Microbiol9:931–941[CrossRef]
    [Google Scholar]
  11. Dramsi S., Biswas I., Maguin E., Braun L., Mastroeni P., Cossart P.. 1995; Entry of Listeria monocytogenes into hepatocytes requires expression of InIB, a surface protein of the internalin multigene family. Mol Microbiol16:251–261[CrossRef]
    [Google Scholar]
  12. Dramsi S., Dehoux P., Lebrun M., Goossens P. L., Cossart P.. 1997; Identification of four new members of the internalin multigene family of Listeria monocytogenes EGD. Infect Immun65:1615–1625
    [Google Scholar]
  13. Drevets D. A., Campbell P. A.. 1991; Roles of complement receptor type 3 in phagocytosis of Listeria monocytogenes by inflammatory mouse peritoneal macrophages. Infect Immun59:2645–2652
    [Google Scholar]
  14. Drevets D. A., Canono B. P., Campbell P. A.. 1992; Listericidal and nonlistericidal mouse macrophages differ in complement receptor type 3-mediated phagocytosis of L. monocytogenes and in preventing escape of the bacteria into the cytoplasm. J Leukocyte Biol52:70–79
    [Google Scholar]
  15. Drevets D. A., Sawyer R. T., Potter T. A., Campbell P. A.. 1995; Listeria monocytogenes infects human endothelial cells by two distinct mechanisms. Infect Immun63:4268–4276
    [Google Scholar]
  16. Dunne D. W., Resnick D., Greenberg J., Krieger M., Joiner K. A.. 1994; The type I macrophage scavenger receptor binds to Gram-positive bacteria and recognizes lipoteichoic acid. Proc Natl Acad Sci USA91:1863–1867[CrossRef]
    [Google Scholar]
  17. Farber J. M., Peterkin P. I.. 1991; Listeria monocytogenes, a food-borne pathogen. Microbiol Rev55:476–511
    [Google Scholar]
  18. Finlay B. B., Cossart P.. 1997; Exploitation of mammalian host cell function by bacterial pathogens. Science276:718–725[CrossRef]
    [Google Scholar]
  19. Gaillard J. L., Finlay B. B.. 1996; Effects of cell polarization and differentiation on entry of Listeria monocytogenes into the enterocyte-like Caco-2 cell line. Infect Immun64:1299–1308
    [Google Scholar]
  20. Gaillard J. L., Berche P., Sansonetti P.. 1986; Transposon mutagenesis as a tool to study the role of hemolysin in the virulence of Listeria monocytogenes. Infect Immun52:50–55
    [Google Scholar]
  21. Gaillard J. L., Berche P., Mounier J., Richard S., Sansonetti P. J.. 1987; In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun55:2822–2829
    [Google Scholar]
  22. Gaillard J. L., Berche P., Fréhel C., Gouin E., Cossart P.. 1991; Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from gram-positive cocci. Cell65:1127–1141[CrossRef]
    [Google Scholar]
  23. Gaillard J. L., Jaubert F., Berche P.. 1996; The inlAB locus mediates the entry of Listeria monocytogenes into hepatocytes in vivo. J Exp Med183:359–369[CrossRef]
    [Google Scholar]
  24. Gellin B. G., Broome C. V., Bibb W. F., Weaver R. E., Gaventa S., Mascola L.. 1991; The epidemiology of listeriosis in the United States – 1986. Listeriosis Study Group. Am J Epidemiol133:392–401
    [Google Scholar]
  25. Gholizadeh Y., Poyart C., Juvin M., Beretti J. L., Croizé J., Berche P., Gaillard J. L.. 1996; Serodiagnosis of listeriosis based upon detection of antibodies against recombinant truncated forms of listeriolysin O. J Clin Microbiol34:1391–1395
    [Google Scholar]
  26. Gray M. L., Killinger A. H.. 1966; Listeria monocytogenes and listeric infections. Bacteriol Rev30:309–382
    [Google Scholar]
  27. Havell E. A.. 1986; Synthesis and secretion of interferon by murine fibroblasts in response to intracellular Listeria monocytogenes. Infect Immun54:787–792
    [Google Scholar]
  28. Heilmann C., Hussain M., Peters G., Gotz F.. 1997; Evidence for autolysin-mediated primary attachment of Staphylococcus epidermidis to a polystyrene surface. Mol Microbiol24:1013–1024[CrossRef]
    [Google Scholar]
  29. Hell W., Meyer H. G., Gatermann S. G.. 1998; Cloning of aas, a gene encoding a Staphylococcus saprophyticus surface protein with adhesive and autolytic properties. Mol Microbiol29:871–881[CrossRef]
    [Google Scholar]
  30. Kaufmann S. H. E.. 1993; Immunity to intracellular bacteria. Annu Rev Immunol11:129–163[CrossRef]
    [Google Scholar]
  31. Kuhn M., Goebel W.. 1989; Identification of an extracellular protein of Listeria monocytogenes possibly involved in intracellular uptake by mammalian cells. Infect Immun57:55–61
    [Google Scholar]
  32. Laemmli U. K.. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227:680–685[CrossRef]
    [Google Scholar]
  33. Mackaness G. B.. 1962; Cellular resistance to infection. J Exp Med116:381–406[CrossRef]
    [Google Scholar]
  34. McLaughlan A. M., Foster S. J.. 1998; Molecular characterization of an autolytic amidase of Listeria monocytogenes EGD. Microbiology144:1359–1367[CrossRef]
    [Google Scholar]
  35. Mengaud J., Ohayon H., Gounon P., Mège R. M., Cossart P.. 1996; E-cadherin is the receptor for internalin, a surface protein required for entry of L. monocytogenes into epithelial cells. Cell84:923–932[CrossRef]
    [Google Scholar]
  36. Nieman R. E., Lorber B.. 1980; Listeriosis in adults: a changing pattern. Report of eight cases and review of the literature 1968–1978. Rev Infect Dis2:207–227[CrossRef]
    [Google Scholar]
  37. Notermans S. H., Dufrenne J., Leimeister-Wachter M., Domann E., Chakraborty T.. 1991; Phosphatidylinositol-specific phospholipase C activity as a marker to distinguish between pathogenic and nonpathogenic Listeria species. Appl Environ Microbiol57:2666–2670
    [Google Scholar]
  38. Pandiripally V. K., Westbrook D. G., Sunki G. R., Bhunia A. K.. 1999; Surface protein p104 is involved in adhesion of Listeria monocytogenes to human intestinal cell line, Caco-2. J Med Microbiol48:117–124[CrossRef]
    [Google Scholar]
  39. Portnoy D. A., Chakraborty T., Goebel W., Cossart P.. 1992; Molecular determinants of Listeria monocytogenes pathogenesis. Infect Immun60:1263–1267
    [Google Scholar]
  40. Poyart-Salmeron C., Trieu-Cuot P., Carlier C., MacGowan A., McLauchlin J., Courvalin P.. 1992; Genetic basis of tetracycline resistance in clinical isolates of Listeria monocytogenes. Antimicrob Agents Chemother36:463–466[CrossRef]
    [Google Scholar]
  41. Rosen H., Gordon S., North R. J.. 1989; Exacerbation of murine listeriosis by a monoclonal antibody specific for the type 3 complement receptor of myelomonocytic cells. Absence of monocytes at infective foci allows Listeria to multiply in nonphagocytic cells. J Exp Med170:27–37[CrossRef]
    [Google Scholar]
  42. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  43. Sheehan B., Kocks C., Dramsi S., Gouin E., Klarsfeld A. D., Mengaud J., Cossart P.. 1994; Molecular and genetic determinants of the Listeria monocytogenes infectious process. Curr Top Microbiol Immunol192:187–216
    [Google Scholar]
  44. Spitzer E. D., Weiss B.. 1985; dfp gene of Escherichia coli K-12, a locus affecting DNA synthesis, codes for a flavoprotein. J Bacteriol164:994–1003
    [Google Scholar]
  45. Spitzer E. D., Jimenez-Billini H. E., Weiss B.. 1988; β-Alanine auxotrophy associated with dfp, a locus affecting DNA synthesis in Escherichia coli. J Bacteriol170:872–876
    [Google Scholar]
  46. Wood S., Maroushek N., Czuprynski C. J.. 1993; Multiplication of Listeria monocytogenes in a murine hepatocyte cell line. Infect Immun61:3068–3072
    [Google Scholar]
  47. Woodcock D. M., Crowther P. J., Doherty J., Jefferson S., DeCruz E., Noyer-Weidner M., Smith S. S., Michael M. Z., Graham M. W.. 1989; Quantitative evaluation of Escherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants. Nucleic Acids Res17:3469–3478[CrossRef]
    [Google Scholar]
  48. Yanisch-Perron C., Vieira J., Messing J.. 1985; Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene33:103–119[CrossRef]
    [Google Scholar]
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