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 .

Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26329-0
2003-06-01
2019-11-12
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/6/mic1491399.html?itemId=/content/journal/micro/10.1099/mic.0.26329-0&mimeType=html&fmt=ahah

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.[CrossRef]
    [Google Scholar]
  2. Arends, J. P. & Zanen, H. C. ( 1988; ). Meningitis caused by Streptococcus suis in humans. Rev Infect Dis 10, 131–137.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [Google Scholar]
  11. Demple, B. ( 1999; ). Radical ideas: genetic responses to oxidative stress. Clin Exp Pharmacol Physiol 26, 64–68.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [Google Scholar]
  26. Miller, J. ( 1972; ). Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  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.[CrossRef]
    [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.[CrossRef]
    [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.
  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 G. von Heijne. Austin: R. G. Landes.
  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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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 Streptococcus 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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [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.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26329-0
Loading
/content/journal/micro/10.1099/mic.0.26329-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error