Journal of Biomedical Science

, Volume 12, Issue 4, pp 587–599 | Cite as

Roles of the Minor Pseudopilins, XpsH, XpsI and XpsJ, in the Formation of XpsG-Containing Pseudopilus in Xanthomonas campestris pv. Campestris

  • Wei-Wen Kuo
  • Hung-Wei Kuo
  • Chun-Chia Cheng
  • Hsiang-Ling Lai
  • Ling-Yun Chen
Article

Summary

Due to their similarity to type IV pilus (Tfp) subunits, the pseudopilins, XpsG, -H, -I, -J and -K, have been predicted to form a pilus-like structure in the type II secretion (T2S) pathway. While overexpression of GspG can result in the formation of bundle structures, the functions of other pseudopilin are not known yet. In this study, we investigate the mutual interaction among the pseudopilins and characterize the specialized minor pseudopilin, XpsJ. By using gel filtration and Ni-NTA affinity chromatography, a linearly ordered interactive relationship is revealed among the four pseudopilins, XpsG-XpsI-XpsH-XpsJ. Notably, unlike the mutant XpsJ194 staying in the inner membrane, wild type XpsJ stayed in the outer membrane and blocked the extension of overexpressed XpsG to outside of the cell. By analogy with the Type I pilus structures, we hypothesize that the XpsH and XpsI might act as an adaptor to connect XpsJ with the major pseudopilin XpsG, and XpsJ might act as a tip to restrict the out-growth of XpsG in the pilus-like structure of the T2S pathway.

Keywords

major pseudopilin minor pseudopilin tip type IV pili type I pili type II secretion pathway 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Possot, O.M., Vignon, G., Bomchil, N., Ebel, F., Pugsley, A.P. (2000) "Multiple interactions between pullulanase secreton components involved in stabilization and cytoplasmic membrane association of PulE" J. Bacteriol. 182: 2142-2152CrossRefPubMedGoogle Scholar
  2. 2.
    Nunn, D.N., Lory, S. (1992) "Components of the protein-excretion apparatus of Pseudomonas aeruginosa are processed by the type IV prepilin peptidase" Proc. Natl. Acad. Sci. USA 89: 47-51PubMedCrossRefGoogle Scholar
  3. 3.
    Nunn, D.N., Lory, S. (1993) "Cleavage, methylation, and localization of the Pseudomonas aeruginosa export proteins XcpT, -U, -V, and -W" J. Bacteriol. 175: 4375-4382PubMedGoogle Scholar
  4. 4.
    Pugsley, A.P., Dupuy, B. (1992) "An enzyme with type IV prepilin peptidase activity is required to process components of the general extracellular protein secretion pathway of Klebsiella oxytoca " Mol. Microbiol. 6: 751-760PubMedCrossRefGoogle Scholar
  5. 5.
    Parge, H.E., Forest, K.T., Hickey, M.J., Christensen, D.A., Getzoff, E.D., Tainer, J.A. (1995) "Structure of the fibre-forming protein pilin at 2.6 A resolution" Nature 378: 32-38CrossRefPubMedGoogle Scholar
  6. 6.
    Keizer, D.W., Slupsky, C.M., Kalisiak, M., Campbell, A.P., Crump, M.P., Sastry, P.A., Hazes, B., Irvin, R.T., Sykes, B.D. (2001) "Structure of a pilin monomer from Pseudomonas aeruginosa: implications for the assembly of pili" J. Biol. Chem. 276: 24186-24193CrossRefPubMedGoogle Scholar
  7. 7.
    Kohler, R., Schafer, K., Muller, S., Vignon, G., Diederichs, K., Philippsen, A., Ringler, P., Pugsley, A.P., Engel, A., Welte, W. (2004) "Structure and assembly of the pseudopilin PulG" Mol. Microbiol. 54: 647-664CrossRefPubMedGoogle Scholar
  8. 8.
    Hobbs, M., Mattick, J.S. (1993) "Common components in the assembly of type 4 fimbriae, DNA transfer systems, filamentous phage and protein-secretion apparatus: a general system for the formation of surface-associated protein complexes" Mol. Microbiol. 10: 233-243PubMedCrossRefGoogle Scholar
  9. 9.
    Pugsley, A.P. (1993) "The complete general secretory pathway in gram-negative bacteria" Microbiol. Rev. 57: 50-108PubMedGoogle Scholar
  10. 10.
    Pugsley, A.P. (1996) "Multimers of the precursor of a type IV pilin-like component of the general secretory pathway are unrelated to pili" Mol. Microbiol. 20: 1235-1245PubMedCrossRefGoogle Scholar
  11. 11.
    Lu, H.M., Motley, S.T., Lory, S. (1997) "Interactions of the components of the general secretion pathway: role of Pseudomonas aeruginosa type IV pilin subunits in complex formation and extracellular protein secretion" Mol. Microbiol. 25: 247-259CrossRefPubMedGoogle Scholar
  12. 12.
    Sauvonnet, N., Vignon, G., Pugsley, A.P., Gounon, P. (2000) "Pilus formation and protein secretion by the same machinery in Escherichia coli " Embo. J. 19: 2221-2228CrossRefPubMedGoogle Scholar
  13. 13.
    Durand, E., Bernadac, A., Ball, G., Lazdunski, A., Sturgis, J.N., Filloux, A. (2003) "Type II protein secretion in Pseudomonas aeruginosa: the pseudopilus is a multifibrillar and adhesive structure" J. Bacteriol. 185: 2749-2758CrossRefPubMedGoogle Scholar
  14. 14.
    Hu, N.T., Leu, W.M., Lee, M.S., Chen, A., Chen, S.C., Song, Y.L., Chen, L.Y. (2002) "XpsG, the major pseudopilin in Xanthomonas campestris pv. campestris, forms a pilus-like structure between cytoplasmic and outer membranes" Biochem. J. 365: 205-211CrossRefPubMedGoogle Scholar
  15. 15.
    Hu, N.T., Hung, M.N., Chiou, S.J., Tang, F., Chiang, D.C., Huang, H.Y., Wu, C.Y. (1992) "Cloning and characterization of a gene required for the secretion of extracellular enzymes across the outer membrane by Xanthomonas campestris pv. campestris" J. Bacteriol. 174: 2679-2687PubMedGoogle Scholar
  16. 16.
    Somogyi, M. (1960) "Modifications of two methods for the assay of amylase" Clin. Chem. 6: 23-35PubMedGoogle Scholar
  17. 17.
    Chen, L.Y., Chen, D.Y., Miaw, J., Hu, N.T. (1996) "XpsD, an outer membrane protein required for protein secretion by Xanthomonas campestris pv. campestris, forms a multimer" J. Biol. Chem. 271: 2703-2708CrossRefPubMedGoogle Scholar
  18. 18.
    Lee, H.M., Wang, K.C., Liu, Y.L., Yew, H.Y., Chen, L.Y., Leu, W.M., Chen, D.C., Hu, N.T. (2000) "Association of the cytoplasmic membrane protein XpsN with the outer membrane protein XpsD in the type II protein secretion apparatus of Xanthomonas campestris pv. campestris" J. Bacteriol. 182: 1549-1557CrossRefPubMedGoogle Scholar
  19. 19.
    Alm, R.A., Mattick, J.S. (1997) "Genes involved in the biogenesis and function of type-4 fimbriae in Pseudomonas aeruginosa " Gene 192: 89-98CrossRefPubMedGoogle Scholar
  20. 20.
    Brinton, C.C., Jr. (1965) "The structure, function, synthesis and genetic control of bacterial pili and a molecular model for DNA and RNA transport in gram negative bacteria" Trans. NY. Acad. Sci. 27: 1003-1054Google Scholar
  21. 21.
    Lindberg, F., Lund, B., Johansson, L., Normark, S. (1987) "Localization of the receptor-binding protein adhesin at the tip of the bacterial pilus" Nature 328: 84-87CrossRefPubMedGoogle Scholar
  22. 22.
    Hahn, E., Wild, P., Hermanns, U., Sebbel, P., Glockshuber, R., Haner, M., Taschner, N., Burkhard, P., Aebi, U., Muller, S.A. (2002) "Exploring the 3D molecular architecture of Escherichia coli type 1 pili" J. Mol. Biol. 323: 845-857CrossRefPubMedGoogle Scholar
  23. 23.
    Craig, L., Pique, M.E., Tainer, J.A. (2004) "Type IV pilus structure and bacterial pathogenicity" Nat. Rev. Microbiol. 2: 363-378CrossRefPubMedGoogle Scholar
  24. 24.
    Ramer, S.W., Schoolnik, G.K., Wu, C.Y., Hwang, J., Schmidt, S.A., Bieber, D. (2002) "The type IV pilus assembly complex: biogenic interactions among the bundle-forming pilus proteins of enteropathogenic Escherichia coli " J. Bacteriol. 184: 3457-3465CrossRefPubMedGoogle Scholar
  25. 25.
    Sandkvist, M. (2001) "Biology of type II secretion" Mol. Microbiol. 40: 271-283CrossRefPubMedGoogle Scholar
  26. 26.
    Bleves, S., Voulhoux, R., Michel, G., Lazdunski, A., Tommassen, J., Filloux, A. (1998) "The secretion apparatus of Pseudomonas aeruginosa: identification of a fifth pseudopilin, XcpX (GspK family)" Mol. Microbiol. 27: 31-40CrossRefPubMedGoogle Scholar

Copyright information

© National Science Council, Taipei 2005

Authors and Affiliations

  • Wei-Wen Kuo
    • 1
  • Hung-Wei Kuo
    • 1
  • Chun-Chia Cheng
    • 1
  • Hsiang-Ling Lai
    • 1
  • Ling-Yun Chen
    • 1
  1. 1.Institute of BiochemistryChung Shan Medical UniversityTaiwanRepublic of China

Personalised recommendations