1887

Microbiology Society logo

Volume 153, Issue 9

Analysis of functional domains present in the N-terminus of the SipB protein Free

Abstract

SipB (593 aa), one of the invasion proteins (Sips), is secreted via the pathogenicity island 1 (SPI-1) type III secretion system (T3SS). Here, we report the delineation of several functional regions present in the SipB protein. Our data show that residues 3–8 of the SipB protein are essential for its secretion from the bacterial cell and that the SicA chaperone, which is important to ensure stability of SipB and SipC in the bacterial cytosol, binds to SipB somewhere between amino acids 80 and100 of the SipB N-terminal region. Interestingly, the N-terminal region (residues 1–160) of SipB (SipB160) cannot be secreted via the SPI-1 T3SS, but fusion of the C-terminal amphipathic region (residues 300–593) to SipB160 can restore secretion via this system.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): FragC, tetanus toxin fragment C , GST, glutathione S-transferase , LLO, listeriolysin O , rSipB, recombinant SipB , Sip, Salmonella invasion protein , SPI, Salmonella pathogenicity island and T3SS, type III secretion system
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.2007/007872-0
2007-09-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/153/9/2998.html?itemId=/content/journal/micro/10.1099/mic.0.2007/007872-0&mimeType=html&fmt=ahah

References

  1. Auvray F., Thomas J., Fraser G. M., Hughes C. 2001; Flagellin polymerisation control by a cytosolic export chaperone. J Mol Biol 308:221–229
     [Google Scholar]
  2. Bennett J. C., Hughes C. 2000; From flagellum assembly to virulence: the extended family of type III export chaperones. Trends Microbiol 8:202–204
     [Google Scholar]
  3. Bennett J. C., Thomas J., Fraser G. M., Hughes C. 2001; Substrate complexes and domain organization of the Salmonella flagellar export chaperones FlgN and FliT. Mol Microbiol 39:781–791
     [Google Scholar]
  4. Birtalan S. C., Phillips R. M., Ghosh P. 2002; Three-dimensional secretion signals in chaperone-effector complexes of bacterial pathogens. Mol Cell 9:971–980
     [Google Scholar]
  5. Bullas L. R., Ryu J. I. 1983; Salmonella typhimurium LT2 strains which are r− m+ for all three chromosomally located systems of DNA restriction and modification. J Bacteriol 156:471–474
     [Google Scholar]
  6. Cheng L. W., Anderson D. M., Schneewind O. 1997; Two independent type III secretion mechanisms for YopE in Yersinia enterocolitica. Mol Microbiol 24:757–765
     [Google Scholar]
  7. Collazo C. M., Galan J. E. 1997; The invasion-associated type III system of Salmonella typhimurium directs the translocation of Sip proteins into the host cell. Mol Microbiol 24:747–756
     [Google Scholar]
  8. Cornelis G. R., Van Gijsegem F. 2000; Assembly and function of type III secretory systems. Annu Rev Microbiol 54:735–774
     [Google Scholar]
  9. Curtiss R. III, Porter S. B., Munson M., Tinge S. A., Hassan J. O., Gentry-Weeks C., Kelly S. M. 1991; Nonrecombinant and recombinant avirulent salmonella vaccines for poultry. In Colonization Control of Human Bacterial Enteropathogens in Poultry pp 169–198 Edited by Blankenship L. C., Bailey J. H. S., Cox N. A., Stern N. J., Meinersmann R. J. New York: Academic Press;
     [Google Scholar]
  10. Darwin K. H., Miller V. L. 2001; Type III secretion chaperone-dependent regulation: activation of virulence genes by SicA and InvF in Salmonella typhimurium. EMBO J 20:1850–1862
     [Google Scholar]
  11. Davis R. W., Bolstein D., Roth J. R. 1980 Advanced Bacterial Genetics Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  12. Eichelberg K., Galán J. E. 2000; The flagellar sigma factor FliA ( σ28) regulates the expression of Salmonella genes associated with the centisome 63 type III secretion system. Infect Immun 68:2735–2743
     [Google Scholar]
  13. Francis M. S., Wolf-Watz H. 1998; YopD of Yersinia pseudotuberculosis is translocated into the cytosol of HeLa epithelial cells: evidence of a structural domain necessary for translocation. Mol Microbiol 29:799–813
     [Google Scholar]
  14. Fraser G. M., Bennett J. C., Hughes C. 1999; Substrate-specific binding of hook-associated proteins by FlgN and FliT, putative chaperones for flagellum assembly. Mol Microbiol 32:569–580
     [Google Scholar]
  15. Fu Y., Galan J. E. 1998; Identification of a specific chaperone for SptP, a substrate of the centisome 63 type III secretion system of Salmonella typhimurium. J Bacteriol 180:3393–3399
     [Google Scholar]
  16. Galán J. E. 2001; Salmonella interactions with host cells: type III secretion at work. Annu Rev Cell Dev Biol 17:53–86
     [Google Scholar]
  17. Galán J. E., Collmer A. 1999; Type III secretion machines: bacterial devices for protein delivery into host cells. Science 284:1322–1328
     [Google Scholar]
  18. Galan J. E., Curtiss R. III 1989; Cloning and molecular characterization of genes whose products allow Salmonella typhimurium to penetrate tissue culture cells. Proc Natl Acad Sci U S A 86:6383–6387
     [Google Scholar]
  19. Galan J. E., Ginocchio C., Costeas P. 1992; Molecular and functional characterization of the Salmonella invasion gene invA: homology of InvA to members of a new protein family. J Bacteriol 174:4338–4349
     [Google Scholar]
  20. Galen J. E., Levine M. M. 2001; Can a ‘flawless’ live vector vaccine strain be engineered?. Trends Microbiol 9:372–376
     [Google Scholar]
  21. Guan K. L., Dixon J. E. 1991; Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem 192:262–267
     [Google Scholar]
  22. Hayward R. D., Koronakis V. 1999; Direct nucleation and bundling of actin by the SipC protein of invasive Salmonella. EMBO J 18:4926–4934
     [Google Scholar]
  23. Hayward R. D., McGhie E. J., Koronakis V. 2000; Membrane fusion activity of purified SipB, a Salmonella surface protein essential for mammalian cell invasion. Mol Microbiol 37:727–739
     [Google Scholar]
  24. Hensel M., Shea J. E., Gleeson C., Jones M. D., Dalton E., Holden D. W. 1995; Simultaneous identification of bacterial virulence genes by negative selection. Science 269:400–403
     [Google Scholar]
  25. Hersh D., Monack D. M., Smith M. R., Ghori N., Falkow S., Zychlinsky A. 1999; The Salmonella invasin SipB induces macrophage apoptosis by binding to caspase-1. Proc Natl Acad Sci U S A 96:2396–2401
     [Google Scholar]
  26. Hueck C. J. 1998; Type III protein secretion systems in bacterial pathogens of animals and plants. Microbiol Mol Biol Rev 62:379–433
     [Google Scholar]
  27. Igwe E. I., Geginat G., Russmann H. 2002; Concomitant cytosolic delivery of two immunodominant listerial antigens by Salmonella enterica serovar Typhimurium confers superior protection against murine listeriosis. Infect Immun 70:7114–7119
     [Google Scholar]
  28. Kimbrough T. G., Miller S. I. 2002; Assembly of the type III secretion needle complex of Salmonella typhimurium. Microbes Infect 4:75–82
     [Google Scholar]
  29. Kubori T., Galan J. E. 2002; Salmonella type III secretion-associated protein InvE controls translocation of effector proteins into host cells. J Bacteriol 184:4699–4708
     [Google Scholar]
  30. Lee S. H., Galan J. E. 2004; Salmonella type III secretion-associated chaperones confer secretion-pathway specificity. Mol Microbiol 51:483–495
     [Google Scholar]
  31. Lloyd S. A., Norman M., Rosqvist R., Wolf-Watz H. 2001; Yersinia YopE is targeted for type III secretion by N-terminal, not mRNA, signals. Mol Microbiol 39:520–531
     [Google Scholar]
  32. McGhie E. J., Hume P. J., Hayward R. D., Torres J., Koronakis V. 2002; Topology of the Salmonella invasion protein SipB in a model bilayer. Mol Microbiol 44:1309–1321
     [Google Scholar]
  33. Michiels T., Cornelis G. R. 1991; Secretion of hybrid proteins by the Yersinia Yop export system. J Bacteriol 173:1677–1685
     [Google Scholar]
  34. Neyt C., Cornelis G. R. 1999; Role of SycD, the chaperone of the Yersinia Yop translocators YopB and YopD. Mol Microbiol 31:143–156
     [Google Scholar]
  35. Ochman H., Soncini F. C., Solomon F., Groisman E. A. 1996; Identification of a pathogenicity island required for Salmonella survival in host cells. Proc Natl Acad Sci U S A 93:7800–7804
     [Google Scholar]
  36. Page A. L., Fromont-Racine M., Sansonetti P., Legrain P., Parsot C. 2001; Characterization of the interaction partners of secreted proteins and chaperones of Shigella flexneri. Mol Microbiol 42:1133–1145
     [Google Scholar]
  37. Parsot C., Hamiaux C., Page A. L. 2003; The various and varying roles of specific chaperones in type III secretion systems. Curr Opin Microbiol 6:7–14
     [Google Scholar]
  38. Ramamurthi K. S., Schneewind O. 2005; A synonymous mutation in Yersinia enterocolitica yopE affects the function of the YopE type III secretion signal. J Bacteriol 187:707–715
     [Google Scholar]
  39. Rüssmann H., Shams H., Poblete F., Fu Y., Galán J. E., Donis R. O. 1998; Delivery of epitopes by the Salmonella type III secretion system for vaccine development. Science 281:565–568
     [Google Scholar]
  40. Rüssmann H., Igwe E. I., Sauer J., Hardt W. D., Bubert A., Geginat G. 2001; Protection against murine listeriosis by oral vaccination with recombinant Salmonella expressing hybrid Yersinia type III proteins. J Immunol 167:357–365
     [Google Scholar]
  41. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  42. Scherer C. A., Cooper E., Miller S. I. 2000; The Salmonella type III secretion translocon protein SspC is inserted into the epithelial cell plasma membrane upon infection. Mol Microbiol 37:1133–1145
     [Google Scholar]
  43. Sory M. P., Boland A., Lambermount I., Cornelis G. 1995; Identification of the YopE and YopH domains required for secretion and internalization into the cytosol of macrophages, using the cyaA gene fusion approach. Proc Natl Acad Sci U S A 92:11998–12002
     [Google Scholar]
  44. Stebbins C. E., Galán J. E. 2001; Maintenance of an unfolded polypeptide by a cognate chaperone in bacterial type III secretion. Nature 414:77–81
     [Google Scholar]
  45. Stecher B., Hapfelmeier S., Muller C., Kremer M., Stallmach T., Hardt W. D. 2004; Flagella and chemotaxis are required for efficient induction of Salmonella enterica serovar Typhimurium colitis in streptomycin-pretreated mice. Infect Immun 72:4138–4150
     [Google Scholar]
  46. Tengel T., Sethson I., Francis M. S. 2002; Conformational analysis by CD and NMR spectroscopy of a peptide encompassing the amphipathic domain of YopD from Yersinia. Eur J Biochem 269:3659–3668
     [Google Scholar]
  47. Tucker S. C., Galan J. E. 2000; Complex function for SicA, a Salmonella enterica serovar Typhimurium type III secretion-associated chaperone. J Bacteriol 182:2262–2268
     [Google Scholar]
  48. Wattiau P., Woestyn S., Cornelis G. R. 1996; Customized secretion chaperones in pathogenic bacteria. Mol Microbiol 20:255–262
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.2007/007872-0
Loading
Analysis of functional domains present in the N-terminus of the SipB protein
Microbiology 153, 2998 (2007); https://doi.org/10.1099/mic.0.2007/007872-0
/content/journal/micro/10.1099/mic.0.2007/007872-0
/content/journal/micro/10.1099/mic.0.2007/007872-0
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