1887

Microbiology Society logo

Volume 159, Issue Pt_11

subsp. DsbA homologue: a thioredoxin-like protein with chaperone function Free

Abstract

is a highly infectious facultative intracellular bacterium and aetiological agent of tularaemia. The conserved hypothetical lipoprotein with homology to thiol/disulphide oxidoreductase proteins (FtDsbA) is an essential virulence factor in . Its protein sequence has two different domains: the DsbA_Com1_like domain (DSBA), with the highly conserved catalytically active site CXXC and -proline residue; and the domain amino-terminal to FKBP-type peptidyl-prolyl isomerases (FKBP_N). To establish the role of both domains in tularaemia infection models, site-directed and deletion mutagenesis affecting the active site (AXXA), the -proline (P286T) and the FKBP_N domain (ΔFKBP_N) were performed. The generated mutations led to high attenuation with the ability to induce full or partial host protective immunity. Recombinant protein analysis revealed that the active site CXXC as well as the -proline residue and the FKBP_N domain are necessary for correct thiol/disulphide oxidoreductase activity. By contrast, only the DSBA domain (and not the FKBP_N domain) seems to be responsible for the chaperone activity of the FtDsbA protein.

  • Received:
  • Accepted:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.070516-0
2013-11-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/159/11/2364.html?itemId=/content/journal/micro/10.1099/mic.0.070516-0&mimeType=html&fmt=ahah

References

  1. Arié J. P., Sassoon N., Betton J. M.( 2001). Chaperone function of FkpA, a heat shock prolyl isomerase, in the periplasm of Escherichia coli. Mol Microbiol 39:199–210 [View Article][PubMed]
     [Google Scholar]
  2. Bessette P. H., Qiu J., Bardwell J. C., Swartz J. R., Georgiou G.( 2001). Effect of sequences of the active-site dipeptides of DsbA and DsbC on in vivo folding of multidisulfide proteins in Escherichia coli. J Bacteriol 183:980–988 [View Article][PubMed]
     [Google Scholar]
  3. Bönquist L., Lindgren H., Golovliov I., Guina T., Sjöstedt A.( 2008). MglA and Igl proteins contribute to the modulation of Francisella tularensis live vaccine strain-containing phagosomes in murine macrophages. Infect Immun 76:3502–3510 [View Article][PubMed]
     [Google Scholar]
  4. Buchner J., Grallert H., Jakob U.( 1998). Analysis of chaperone function using citrate synthase as nonnative substrate protein. Methods Enzymol 290:323–338 [View Article][PubMed]
     [Google Scholar]
  5. Burall L. S., Harro J. M., Li X., Lockatell C. V., Himpsl S. D., Hebel J. R., Johnson D. E., Mobley H. L. T.( 2004). Proteus mirabilis genes that contribute to pathogenesis of urinary tract infection: identification of 25 signature-tagged mutants attenuated at least 100-fold. Infect Immun 72:2922–2938 [View Article][PubMed]
     [Google Scholar]
  6. Celli J.( 2008). Intracellular localization of Brucella abortus and Francisella tularensis in primary murine macrophages. Methods Mol Biol 431:133–145[PubMed]
     [Google Scholar]
  7. Chamberlain R. E.( 1965). Evaluation of live tularemia vaccine prepared in a chemically defined medium. Appl Microbiol 13:232–235[PubMed]
     [Google Scholar]
  8. Charbonnier J. B., Belin P., Moutiez M., Stura E. A., Quéméneur E.( 1999). On the role of the cis-proline residue in the active site of DsbA. Protein Sci 8:96–105 [View Article][PubMed]
     [Google Scholar]
  9. Chen J., Song J. L., Zhang S., Wang Y., Cui D. F., Wang C. C.( 1999). Chaperone activity of DsbC. J Biol Chem 274:19601–19605 [View Article][PubMed]
     [Google Scholar]
  10. Chivers P. T., Prehoda K. E., Raines R. T.( 1997). The CXXC motif: a rheostat in the active site. Biochemistry 36:4061–4066 [View Article][PubMed]
     [Google Scholar]
  11. Ha U.-H., Wang Y., Jin S.( 2003). DsbA of Pseudomonas aeruginosa is essential for multiple virulence factors. Infect Immun 71:1590–1595 [View Article][PubMed]
     [Google Scholar]
  12. Hendrix L. R., Mallavia L. P., Samuel J. E.( 1993). Cloning and sequencing of Coxiella burnetii outer membrane protein gene com1. Infect Immun 61:470–477[PubMed]
     [Google Scholar]
  13. Heras B., Totsika M., Jarrott R., Shouldice S. R., Guncar G., Achard M. E. S., Wells T. J., Argente M. P., McEwan A. G., Schembri M. A.( 2010). Structural and functional characterization of three DsbA paralogues from Salmonella enterica serovar Typhimurium. J Biol Chem 285:18423–18432 [View Article][PubMed]
     [Google Scholar]
  14. Holmgren A.( 1979). Thioredoxin catalyzes the reduction of insulin disulfides by dithiothreitol and dihydrolipoamide. J Biol Chem 254:9627–9632[PubMed]
     [Google Scholar]
  15. Jameson-Lee M., Garduño R. A., Hoffman P. S.( 2011). DsbA2 (27 kDa Com1-like protein) of Legionella pneumophila catalyses extracytoplasmic disulphide-bond formation in proteins including the Dot/Icm type IV secretion system. Mol Microbiol 80:835–852 [View Article][PubMed]
     [Google Scholar]
  16. Jänsch L., Kruft V., Schmitz U. K., Braun H. P.( 1996). New insights into the composition, molecular mass and stoichiometry of the protein complexes of plant mitochondria. Plant J 9:357–368 [View Article][PubMed]
     [Google Scholar]
  17. Kadokura H., Tian H., Zander T., Bardwell J. C. A., Beckwith J.( 2004). Snapshots of DsbA in action: detection of proteins in the process of oxidative folding. Science 303:534–537 [View Article][PubMed]
     [Google Scholar]
  18. Köhler R., Fanghänel J., König B., Lüneberg E., Frosch M., Rahfeld J.-U., Hilgenfeld R., Fischer G., Hacker J., Steinert M.( 2003). Biochemical and functional analyses of the Mip protein: influence of the N-terminal half and of peptidylprolyl isomerase activity on the virulence of Legionella pneumophila. Infect Immun 71:4389–4397 [View Article][PubMed]
     [Google Scholar]
  19. Lafaye C., Iwema T., Carpentier P., Jullian-Binard C., Kroll J. S., Collet J.-F., Serre L.( 2009). Biochemical and structural study of the homologues of the thiol-disulfide oxidoreductase DsbA in Neisseria meningitidis. J Mol Biol 392:952–966 [View Article][PubMed]
     [Google Scholar]
  20. Liu X., Wang C. C.( 2001). Disulfide-dependent folding and export of Escherichia coli DsbC. J Biol Chem 276:1146–1151 [View Article][PubMed]
     [Google Scholar]
  21. Martin J. L.( 1995). Thioredoxin—a fold for all reasons. Structure 3:245–250 [View Article][PubMed]
     [Google Scholar]
  22. Miki T., Okada N., Danbara H.( 2004). Two periplasmic disulfide oxidoreductases, DsbA and SrgA, target outer membrane protein SpiA, a component of the Salmonella pathogenicity island 2 type III secretion system. J Biol Chem 279:34631–34642 [View Article][PubMed]
     [Google Scholar]
  23. Qin A., Scott D. W., Thompson J. A., Mann B. J.( 2009). Identification of an essential Francisella tularensis subsp. tularensis virulence factor. Infect Immun 77:152–161 [View Article][PubMed]
     [Google Scholar]
  24. Qin A., Scott D. W., Rabideau M. M., Moore E. A., Mann B. J.( 2011). Requirement of the CXXC motif of novel Francisella infectivity potentiator protein B FipB, and FipA in virulence of F. tularensis subsp. tularensis. PLoS ONE 6:e24611 [View Article][PubMed]
     [Google Scholar]
  25. Riboldi-Tunnicliffe A., König B., Jessen S., Weiss M. S., Rahfeld J., Hacker J., Fischer G., Hilgenfeld R.( 2001). Crystal structure of Mip, a prolylisomerase from Legionella pneumophila. Nat Struct Biol 8:779–783 [View Article][PubMed]
     [Google Scholar]
  26. Rodriguez S. A., Yu J.-J., Davis G., Arulanandam B. P., Klose K. E.( 2008). Targeted inactivation of Francisella tularensis genes by group II introns. Appl Environ Microbiol 74:2619–2626 [View Article][PubMed]
     [Google Scholar]
  27. Saul F. A., Arié J.-P., Vulliez-le Normand B., Kahn R., Betton J.-M., Bentley G. A.( 2004). Structural and functional studies of FkpA from Escherichia coli, a cis/trans peptidyl-prolyl isomerase with chaperone activity. J Mol Biol 335:595–608 [View Article][PubMed]
     [Google Scholar]
  28. Schägger H., von Jagow G.( 1991). Blue native electrophoresis for isolation of membrane protein complexes in enzymatically active form. Anal Biochem 199:223–231 [View Article][PubMed]
     [Google Scholar]
  29. Shao F., Bader M. W., Jakob U., Bardwell J. C.( 2000). DsbG, a protein disulfide isomerase with chaperone activity. J Biol Chem 275:13349–13352 [View Article][PubMed]
     [Google Scholar]
  30. Stenson T. H., Weiss A. A.( 2002). DsbA and DsbC are required for secretion of pertussis toxin by Bordetella pertussis. Infect Immun 70:2297–2303 [View Article][PubMed]
     [Google Scholar]
  31. Straskova A., Pavkova I., Link M., Forslund A.-L., Kuoppa K., Noppa L., Kroca M., Fucikova A., Klimentova J.& other authors ( 2009). Proteome analysis of an attenuated Francisella tularensis dsbA mutant: identification of potential DsbA substrate proteins. J Proteome Res 8:5336–5346 [View Article][PubMed]
     [Google Scholar]
  32. Thakran S., Li H., Lavine C. L., Miller M. A., Bina J. E., Bina X. R., Re F.( 2008). Identification of Francisella tularensis lipoproteins that stimulate the toll-like receptor (TLR) 2/TLR1 heterodimer. J Biol Chem 283:3751–3760 [View Article][PubMed]
     [Google Scholar]
  33. Thomas R. M., Twine S. M., Fulton K. M., Tessier L., Kilmury S. L. N., Ding W., Harmer N., Michell S. L., Oyston P. C. F.& other authors ( 2011). Glycosylation of DsbA in Francisella tularensis subsp. tularensis. J Bacteriol 193:5498–5509 [View Article][PubMed]
     [Google Scholar]
  34. Wang C. C., Tsou C. L.( 1993). Protein disulfide isomerase is both an enzyme and a chaperone. FASEB J 7:1515–1517[PubMed]
     [Google Scholar]
  35. Yu J., Oragui E. E., Stephens A., Kroll J. S., Venkatesan M. M.( 2001). Inactivation of DsbA alters the behaviour of Shigella flexneri towards murine and human-derived macrophage-like cells. FEMS Microbiol Lett 204:81–88 [View Article][PubMed]
     [Google Scholar]
  36. Zheng W. D., Quan H., Song J. L., Yang S. L., Wang C. C.( 1997). Does DsbA have chaperone-like activity?. Arch Biochem Biophys 337:326–331 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.070516-0
Loading
Francisella tularensis subsp. holarctica DsbA homologue: a thioredoxin-like protein with chaperone function
Microbiology 159, 2364 (2013); https://doi.org/10.1099/mic.0.070516-0
/content/journal/micro/10.1099/mic.0.070516-0
/content/journal/micro/10.1099/mic.0.070516-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

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