Functional characterization of the Erwinia chrysanthemi Outs protein, an element of a type II secretion system Free

Abstract

SUMMARY: Secretion of pectate lyases and a cellulase occurs in winis chrysanthemi through a type II secretion machinery, the Out system. Proper insertion of the secretin OutD in the outer membrane requires the presence of Outs. Outs is an outer-membrane lipoprotein that interacts directly with OutD. Using ligand-blotting experiments, it has been shown that this interaction requires at least the 62 C-terminal amino acids of OutD. When this domain was added to the C-terminal extremity of the secreted pectate lyase PelD, the construct was stabilized by Outs but not inserted into the outer membrane. Thus, this domain is sufficient to interact with Outs but it is unable to confer the ability to be inserted into the outer membrane in the presence of Outs. A screen for outs mutants unable to secrete pectate lyases gave only mutants unable to properly localize OutD in the outer membrane and no mutant in the protection function. Thus, the interaction between Outs and OutD can probably not be abolished by the mutation of a single amino acid, and the insertion of OutD in the outer membrane may require additional proteins.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-144-11-3219
1998-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/144/11/mic-144-11-3219.html?itemId=/content/journal/micro/10.1099/00221287-144-11-3219&mimeType=html&fmt=ahah

References

  1. Bergler H., Abraham D., Aschauer H., Turnowsky F. 1994; Inhibition of lipid biosynthesis induces the expression of the pspA gene.. Microbiology 140:1937–1944
    [Google Scholar]
  2. Brissette i.L., Russel M. 1990; Secretion and membrane integration of a filamentous phage-encoded morphogenetic protein.. J Mol Biol 211:565–580
    [Google Scholar]
  3. Chen R., Henning U. 1996; A periplasmic protein (Skp) of Escherichia coli selectively binds a class of outer membrane proteins.. Mol Microbiol 19:1287–1294
    [Google Scholar]
  4. Chen L.-Y., Chen D.-Y., Miaw J., Hu N.-T. 1996; XcpD, an outer membrane protein required for protein secretion by Xanthomonas campestris pv.campestris, forms a multimer.. J Mol Biol 271:2703–2708
    [Google Scholar]
  5. Condemine G., Dorel C., Hugouvieux-Cotte-Pattat N., Robert-Baudouy J. 1992; Some of the out genes involved in the secretion of pectate lyases in Erwinia chrysanthemi are regulated by kdgR. . Mol Microbiol 6:3199–3211
    [Google Scholar]
  6. Daefler S., Guilvout I., Hardie K.R., Pugsley A.P., Russel M. 1997a; The C-terminal domain of the secretin PulD contains the binding site for its cognate chaperone, PulS, and confers PulS dependence on pIVfl function.. Mol Microbiol 24:465–475
    [Google Scholar]
  7. Daefler S., Russel M., Model P. 1997b; Module swaps between related translocator proteins pIVfl, pIVIke and PulD: identification of a specificity domain.. J Mol Biol 266:978–992
    [Google Scholar]
  8. Drake S.L., Koomey M. 1995; The product of the pilQ gene is essential for the biogenesis of the type IV pili in Neisseria gonorrhoeae. . Mol Microbiol 18:975–986
    [Google Scholar]
  9. Drake S.L., Sandstedt S.A., Koomey M. 1997; PilP, a pilus biogenesis lipoprotein in Neisseria gonorrhoeae, affects expression of PilQ as a high-molecular-mass multimer.. Mol Microbiol 23:657–668
    [Google Scholar]
  10. Dykxhoorn D.M., St Pierre R., Furlong C.E. 1996; A set of compatible tac promoter expression vectors.. Gene 177:133–136
    [Google Scholar]
  11. d’Enfert C., Pugsley A.P. 1987; Klebsiella pneumoniae pulS gene encodes an outer membrane lipoprotein required for pullulanase secretion.. J Bacteriol 171:3673–3679
    [Google Scholar]
  12. Genin S., Boucher C.A. 1994; A superfamily of proteins involved in different secretion pathways in Gram-negative bacteria : modular structure and specificity of the N-terminal domain.. Mol Gen Genet 243:112–118
    [Google Scholar]
  13. Hardie K.R., Lory S., Pugsley A.P. 1996a; Insertion of an outer membrane protein in Escherichia coli requires a chaperonelike protein.. EMBO J 15:978–988
    [Google Scholar]
  14. Hardie K.R., Seydel A., Guilvout I., Pugsley A.P. 1996b; The secretin-specific, chaperone-like protein of the general secretory pathway: separation of proteolytic protection and piloting functions.. Mol Microbiol 22:967–976
    [Google Scholar]
  15. Hugouvieux-Cotte-Pattat N., Dominguez H., Robert-Baudouy J. 1992; Environmental conditions affect transcription of the pectinase genes of Erwinia chrysanthemi 3937.. J Bacteriol 174:7807–7818
    [Google Scholar]
  16. Johnson B.H., Hecht M.H. 1994; Recombinant proteins can be isolated from E. coli cells by repeated cycles of freezing and thawing.. Bio/Technology 12:1357–1360
    [Google Scholar]
  17. Kleerebezem M., Crielaard W., Tommassen J. 1996; Involvement of stress protein PspA (phage shock protein A) of Escherichia coli in maintenance of the proton motive force under stress conditions.. EMBO J 15:162–171
    [Google Scholar]
  18. Koster M., Bitter W., de Cock H., Allaoui A., Cornelis G.R., Tommassen J. 1997; The outer membrane component, YscC, of the Yop secretion machinery of Yersinia enterocolitica forms a ring-shaped multimeric complex.. Mol Microbiol 26:789–797
    [Google Scholar]
  19. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4.. Nature 227:680–685
    [Google Scholar]
  20. Linderoth N.A., Simon M.A., Russel M. 1997; The filamentous phage pIV multimer visualized by scanning transmission electon microscopy.. Science 278:1635–1638
    [Google Scholar]
  21. de Lorenzo V., Eltis L., Kessler B., Timmis K.N. 1993; Analysis of Pseudomonas gene products using laclq/Ptrp-lac plasmids and transposons that confer conditional phenotypes.. Gene 123:17–24
    [Google Scholar]
  22. Matsuyama S.-l., Tajima T., Tokuda H. 1995; A novel periplasmic carrier protein involved in the sorting and transport of Escherichia coli lipoproteins destined for the outer membrane.. EMBO J 14:3365–3372
    [Google Scholar]
  23. Miller J.H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  24. Missiakas D., Betton J.-M., Raina S. 1997; New components of protein folding in extracytoplasmic compartments of Escherichia coli SurA, FkpA, and Skp/OmpH.. Mol Microbiol 21:871–884
    [Google Scholar]
  25. Model P., Jovanovic G., Dworkin J. 1997; The Escherichia coli phage-shock-protein (psp) operon.. Mol Microbiol 24:255–261
    [Google Scholar]
  26. Moran F., Nasuno S., Starr M.P. 1968; Extracellular and intracellular polygalacturonic transeliminases of Erwinia caro- tovora. . Arch Biochem Biophys 123:298–306
    [Google Scholar]
  27. Noonan B., Trust T.J. 1995; Molecular analysis of an A- protein secretion mutant of Aeromonas salmonicida reveals a surface layer-specific protein secretion pathway.. J Mol Biol 248:316–327
    [Google Scholar]
  28. Roine E., Nunn D.N., Paulin L., Romantschuk M. 1996; Characterization of genes required for pilus expression in Pseudomonas syringae pathovar phaseolicola. . J Bacteriol 178:410–417
    [Google Scholar]
  29. Russel M., Kazmierczak B. 1993; Analysis of the structure of the subcellular location of filamentous phage plV.. J Bacteriol 175:3998–4007
    [Google Scholar]
  30. Russel M., Model P. 1984; Replacement of the ftp gene of Escherichia coli by an inactive gene cloned on a plasmid.. J Bacteriol 159:1034–1039
    [Google Scholar]
  31. 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]
  32. Shevchik V.E., Condemine G., Hugouvieux-Cotte-Pattat N., Robert-Baudouy J. 1996; Characterization of pectin methyl-esterase B, an outer membrane lipoprotein of Erwinia chrys- anthemi 3937.. Mol Microbiol 19:455–466
    [Google Scholar]
  33. Shevchik V.E., Robert-Baudouy J., Condemine G. 1997; Specific interaction between OutD, an Erwinia chrysanthemi outer membrane protein of the general secretory pathway, and secreted proteins.. EMBO J 16:3007–3016
    [Google Scholar]
  34. Tabor S., Richardson C. 1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes.. Proc Natl Acad Sci USA 821074–1078
    [Google Scholar]
  35. Yamaguchi K., Yu F., Inouye M. 1988; A single amino acid determinant of the membrane localization of lipoproteins in E. coli. . Cell 53:423–132
    [Google Scholar]
  36. Yang J., Ganesan S., Sarsero J., Pittard A.J. 1993; A genetic analysis of various functions of the TyrR protein of Escherichia coli. . J Bacteriol 175:1767–1776
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-144-11-3219
Loading
/content/journal/micro/10.1099/00221287-144-11-3219
Loading

Data & Media loading...

Most cited Most Cited RSS feed