1887

Abstract

This paper reports the complete coding sequence for a proliprotein signal peptidase (SP-ase) of , Lsp. This is believed to be the first SP-ase described for . SP-ase II is involved in the removal of the signal peptide from glyceride-modified prolipoproteins. By using transcription/translation systems, it was shown that the gene was transcribed . Functionality of Lsp in was demonstrated by using an globomycin resistance assay, to show that expression of Lsp in increased the globomycin resistance. An isogenic mutant of serotype 2 unable to produce Lsp was constructed and shown to process lipoproteins incorrectly, including an homologue of the pneumococcal PsaA lipoprotein. Five piglets were inoculated with a mixture of both strains in an experimental infection, to determine the virulence of the mutant strain relative to that of the wild-type strain in a competitive challenge experiment. The data showed that both strains were equally virulent, indicating that the knockout mutant of is not attenuated .

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2003-06-01
2024-10-12
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References

  1. Altschul S. F., Madden T. L., Schaffer A. A., Zhang J. H., Zhang Z., Miller W., Lipman D. J. 1997; Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
    [Google Scholar]
  2. Arends J. P., Zanen H. C. 1988; Meningitis caused by Streptococcus suis in humans. Rev Infect Dis 10:131–137
    [Google Scholar]
  3. Berry A. M., Paton J. C. 1996; Sequence heterogeneity of PsaA, a 37-kilodalton putative adhesin essential for virulence of Streptococcus pneumoniae . Infect Immun 64:5255–5262
    [Google Scholar]
  4. Bolotin A., Wincker P., Mauger S., Jaillon O., Malarme K., Weissenbach J., Ehrlich S. D., Sorokin A. 2001; The complete genome sequence of the lactic acid bacterium Lactococcus lactis ssp. lactis IL1403. Genome Res 11:731–753
    [Google Scholar]
  5. Brown J. S., Gilliland S. M., Holden D. W. 2001; A Streptococcus pneumoniae pathogenicity island encoding an ABC transporter involved in iron uptake and virulence. Mol Microbiol 40:572–585
    [Google Scholar]
  6. Burnette-Curley D., Wells V., Viscount H., Munro C. L., Fenno J. C., Fives-Taylor P., Macrina F. L. 1995; FimA, a major virulence factor associated with Streptococcus sanguis endocarditis. Infect Immun 63:4669–4674
    [Google Scholar]
  7. Clifton-Hadley F. A. 1983; Streptococcus suis type 2 infections. Br Vet J 139:1–5
    [Google Scholar]
  8. Conrad J., Sun D., Englund N., Ofengand J. 1998; The rluC gene of Escherichia coli codes for a pseudouridine synthase that is solely responsible for synthesis of pseudouridine at positions 955, 2504, and 2580 in 23S ribosomal RNA. J Biol Chem 273:18562–18566
    [Google Scholar]
  9. Coulter S. N., Schwan W. R., Ng E. Y. W. 7 other authors 1998; Staphylococcus aureus genetic loci impacting growth and survival in multiple infection models. Mol Microbiol 30:393–404
    [Google Scholar]
  10. de Greeff A, Buys H., Verhaar R., Dijkstra J., van Alphen L., Smith H. E. 2002; Contribution of fibronectin-binding protein in pathogenesis of Streptococcus suis serotype 2. Infect Immun 70:1319–1325
    [Google Scholar]
  11. Demple B. 1999; Radical ideas: genetic responses to oxidative stress. Clin Exp Pharmacol Physiol 26:64–68
    [Google Scholar]
  12. Fuller T. E., Martin S., Teel J. F., Alaniz G. R., Kennedy M. J., Lowery D. E. 2000; Identification of Actinobacillus pleuropneumoniae virulence genes using signature-tagged mutagenesis in a swine infection model. Microb Pathog 29:39–51
    [Google Scholar]
  13. Gan K., Gupta S. D., Sankaran K., Schmidt M. B., Wu H. C. 1993; Isolation and characterization of a temperature-sensitive mutant of Salmonella typhimurium defective in prolipoprotein modification. J Biol Chem 268:16544–16550
    [Google Scholar]
  14. Gupta S. D., Gan K., Schmidt M. B., Wu H. C. 1993; Characterization of a temperature-sensitive mutant of Salmonella typhimurium defective in apolipoprotein N- acyltransferase. J Biol Chem 268:16551–16556
    [Google Scholar]
  15. Hamilton A., Harrington D., Sutcliffe I. C. 2000; Characterisation of acid phosphatase activities in the equine pathogen Streptococcus equi . Syst Appl Microbiol 23:325–329
    [Google Scholar]
  16. Harrington D. J., Greated J. S., Chanter N., Sutcliffe I. C. 2000; Identification of lipoprotein homologues of pneumococcal PsaA in the equine pathogens Streptococcus equi and Streptococcus zooepidemicus . Infect Immun 68:6048–6051
    [Google Scholar]
  17. Horinouchi S., Weisblum B. 1982a; Nucleotide sequence and functional map of pE194, a plasmid that specifies inducible resistance to macrolide., lincosamide. J Bacteriol 150:804–814
    [Google Scholar]
  18. Horinouchi S., Weisblum B. 1982b; Nucleotide sequence and functional map of pC194. a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol 150:815–825
    [Google Scholar]
  19. Inukai M., Takeuchi M., Shimizu K., Arai M. 1978; Mechanism of action of globomycin. J Antibiot 31:1203–1205
    [Google Scholar]
  20. Kitten T., Munro C. L., Michalek S. M., Macrina F. L. 2000; Genetic characterisation of a Streptococcus mutans LraI family operon and its role in virulence. Infect Immun 68:4441–4451
    [Google Scholar]
  21. Konings R. N. H., Verhoeven E. J. M., Peeters B. P. H. 1987; pKUN vectors for the separate production of both DNA strands of recombinant plasmids. Methods Enzymol 153:12–34
    [Google Scholar]
  22. Koskiniemi S., Sellin M., Norgren M. 1998; Identification of two genes, cpsX and cpsY , with putative regulatory function on capsule expression in group B streptococci. FEMS Immunol Med Microbiol 21:159–168
    [Google Scholar]
  23. Mahan M. J., Slauch J. M., Mekalanos J. J. 1993; Selection of bacterial virulence genes that are specifically induced in host tissues. Science 259:686–688
    [Google Scholar]
  24. Marra A., Lawson S., Asundi J. S., Brigham D., Hromockyj A. E. 2002; In vivo characterisation of the psa genes from Streptococcus pneumoniae in multiple models of infection. Microbiology 148:1483–1491
    [Google Scholar]
  25. Mei J.-M., Nourbakhsh F., Ford C. W., Holden D. W. 1997; Identification of Staphylococcus aureus virulence genes in a murine model of bacteraemia using signature-tagged mutagenesis. Mol Microbiol 26:399–407
    [Google Scholar]
  26. Miller J. 1972 Experiments in Molecular Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  27. Petit C. M., Brown J. R., Ingraham K., Bryant A. P., Holmes D. J. 2001; Lipid modification of prelipoproteins is dispensable for growth in vitro but essential for virulence in Streptococcus pneumoniae . FEMS Microbiol Lett 200:229–233
    [Google Scholar]
  28. Prágai Z, Tjalsma H., Bolhuis A., van Dijl J. M., Venema G., Bron S. 1997; The signal peptidase II ( lsp) gene of Bacillus subtilis . Microbiology 143:1327–1333
    [Google Scholar]
  29. 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]
  30. Sankaran K., Wu H. C. 1994a; Lipid modification of bacterial prolipoprotein – transfer of diacylglycerol moiety from phosphatidylglycerol. J Biol Chem 269:19701–19706
    [Google Scholar]
  31. Sankaran K., Wu H. C. 1994b; Signal peptidase II – specific signal peptidase for bacterial lipoproteins. In Signal Peptidases pp  17–29 Edited by von Heijne G. Austin: R. G. Landes;
    [Google Scholar]
  32. Smith H. E., Vecht U., Gielkens A. L. J., Smits M. A. 1992; Cloning and nucleotide sequence of the gene encoding the 136-kilodalton surface protein (muramidase-released protein) of Streptococcus suis type 2. Infect Immun 60:2361–2367
    [Google Scholar]
  33. Smith H. E., Wisselink H. J., Vecht U., Gielkens A. L. J., Smits M. A. 1995; High-efficiency transformation and gene inactivation in Streptococcus suis type 2. Microbiology 141:181–188
    [Google Scholar]
  34. Smith H. E., Damman M., van der Velde J., Wagenaar F., Wisselink H. J., Stockhofe-Zurwieden N., Smits M. A. 1999a; Identification and characterization of the cps locus of Streptococcus suis serotype 2: the capsule protects against phagocytosis and is an important virulence factor. Infect Immun 67:1750–1756
    [Google Scholar]
  35. Smith H. E., Veenbergen V., van der Velde J., Damman M., Wisselink H. J., Smits M. A. 1999b; The cps genes of Streptococcus suis serotypes 1, 2 and 9: development of rapid serotype-specific PCR assays. J Clin Microbiol 37:3146–3152
    [Google Scholar]
  36. Smith H. E., Buijs H., de Vries R., Wisselink H. J., Stockhofe-Zurwieden N., Smits M. A. 2001; Environmentally regulated genes of Streptococcus suis : identification by the use of iron-restricted conditions in vitro and by experimental infection of piglets. Microbiology 147:271–280
    [Google Scholar]
  37. Sutcliffe I. C., Harrington D. J. 2002; Pattern searches for the identification of putative lipoprotein genes in Gram-positive bacterial genomes. Microbiology 148:2065–2077
    [Google Scholar]
  38. Sutcliffe I. C., Russell R. R. B. 1995; Lipoproteins of gram-positive bacteria. J Bacteriol 177:1123–1128
    [Google Scholar]
  39. Tettelin H., Nelson K. E., Paulsen I. T. 36 other authors 2001; Complete genome sequence of a virulent isolate of Streptococcus pneumoniae . Science 293:498–506
    [Google Scholar]
  40. Tjalsma H., Kontinen V. P., Prágai Z., Wu H., Meima T., Venema G., Bron S., Sarvas M., van Dijl J. M.. 1999a; The role of lipoprotein processing by signal peptidase II in the Gram-positive eubacterium Bacillus subtilis . J Biol Chem 274:1698–1707
    [Google Scholar]
  41. Tjalsma H., Zanen G., Venema G., Bron S., van Dijl J. M. 1999b; The potential active site of the lipoprotein-specific (type II) signal peptidase of Bacillus subtilis . J Biol Chem 274:28191–28197
    [Google Scholar]
  42. Tokunaga M., Loranger J. M., Wu H. C. 1983; Isolation and characterization of an Escherichia coli clone overproducing prolipoprotein signal peptidase. J Biol Chem 258:12102–12105
    [Google Scholar]
  43. Vecht U., van Leengoed L. A. M. G., Verheyen E. R. M. 1985; Streptococcus suis infections in pigs in The Netherlands (part I. Vet Q 7:315–321
    [Google Scholar]
  44. Vecht U., Arends J. P., van der Molen E. J., van Leengoed L. A. M. G. 1989; Differences in virulence between two strains of S treptococcus suis type 2 after experimentally induced infection of newborn germfree pigs. Am J Vet Res 50:1037–1043
    [Google Scholar]
  45. Vecht U., Wisselink H. J., van Dijk J. E., Smith H. E. 1992; Virulence of Streptococcus suis type 2 strains in newborn germfree pigs depend on phenotype. Infect Immun 60:550–556
    [Google Scholar]
  46. Williams M. G., Fortson M., Dykstra C. C., Jensen P., Kushner S. R. 1989; Identification and genetic mapping of the structural gene for an essential Escherichia coli membrane protein. J Bacteriol 171:565–568
    [Google Scholar]
  47. Wisselink H. J., Reek F. H., Vecht U., Stockhofe-Zurwieden N., Smits M. A., Smith H. E. 1999; Detection of virulent strains of Streptococcus suis type 2 and highly virulent strains of Streptococcus suis type 1 in tonsillar specimens of pigs by PCR. Vet Microbiol 67:143–157
    [Google Scholar]
  48. Wisselink H. J., Joosten J. J., Smith H. E. 2002; Multiplex PCR assays for simultaneous detection of six major serotypes and two virulence-associated phenotypes of Streptococcus suis in tonsillar specimens of pigs. J Clin Microbiol 40:2922–2929
    [Google Scholar]
  49. Witke C., Götz F. 1995; Cloning and nucleotide sequence of the signal peptidase II ( lsp) -gene from Staphylococcus carnosus . FEMS Microbiol Lett 126:233–240
    [Google Scholar]
  50. Yamagata H., Ippolito C., Inukai M., Inouye M. 1982; Temperature-sensitive processing of outer membrane lipoprotein in an Escherichia coli mutant. J Baceteriol 152:1163–1168
    [Google Scholar]
  51. Zhao X.-J., Wu H. C. 1992; Nucleotide sequence of the Staphylococcus aureus signal peptidase II ( lsp) gene. FEBS Lett 299:80–84
    [Google Scholar]
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