1887

Microbiology Society logo

Volume 149, Issue 9

Dissection of homologous translocon operons reveals a distinct role for YopD in type III secretion by Free

Abstract

The homologous and translocase operons of and pathogenic spp., respectively, are responsible for the translocation of anti-host effectors into the cytosol of infected eukaryotic cells. In , this operon is also required for -regulatory control. To probe for key molecular interactions during the infection process, the functional interchangeability of / and / was investigated. Secretion of PopB produced in a Δ null mutant of was only observed when co-produced with its native chaperone PcrH, but this was sufficient to complement the translocation defect. The Δ null mutant synthesized and secreted PopD even in the absence of native PcrH, yet this did not restore YopD-dependent -regulatory control or effector translocation. Thus, this suggests that key residues in YopD, which are not conserved in PopD, are essential for functional type III secretion.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): TTSS, type III secretion system
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26322-0
2003-09-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/micro/149/9/mic1492615.html?itemId=/content/journal/micro/10.1099/mic.0.26322-0&mimeType=html&fmt=ahah

References

  1. Allmond L. R., Karaca T. J., Nguyen V. N., Nguyen T., Wiener-Kronish J. P., Sawa T. 2003; Protein binding between PcrG–PcrV and PcrH–PopB/PopD encoded by the pcrGVH – popBD operon of the Pseudomonas aeruginosa type III secretion system. Infect Immun 71:2230–2233
     [Google Scholar]
  2. Anderson D. M., Ramamurthi K. S., Tam C., Schneewind O. 2002; YopD and LcrH regulate expression of Yersinia enterocolitica YopQ by a posttranscriptional mechanism and bind to yopQ RNA. J Bacteriol 184:1287–1295
     [Google Scholar]
  3. Barbieri J. T. 2000; Pseudomonas aeruginosa exoenzyme S, a bifunctional type-III secreted cytotoxin. Int J Med Microbiol 290:381–387
     [Google Scholar]
  4. Bergman T., Håkansson S., Forsberg Å., Norlander L., Macellaro A., Bäckman A., Bölin I., Wolf-Watz H. 1991; Analysis of the V antigen lcrGVH – yopBD operon of Yersinia pseudotuberculosis : evidence for a regulatory role of LcrH and LcrV. J Bacteriol 173:1607–1616
     [Google Scholar]
  5. Bliska J. B. 2000; Yop effectors of Yersinia spp. and actin rearrangements. Trends Microbiol 8:205–208
     [Google Scholar]
  6. Blocker A., Gounon P., Larquet E., Niebuhr K., Cabiaux V., Parsot C., Sansonetti P. 1999; The tripartite type III secreton of Shigella flexneri inserts IpaB and IpaC into host membranes. J Cell Biol 147:683–693
     [Google Scholar]
  7. Bölin I., Wolf-Watz H. 1984; Molecular cloning of the temperature-inducible outer membrane protein 1 of Yersinia pseudotuberculosis . Infect Immun 43:72–78
     [Google Scholar]
  8. Bröms J. E., Sundin C., Francis M. S., Forsberg Å. 2003a; Comparative analysis of type III effector translocation by Yersinia pseudotuberculosis expressing native LcrV or PcrV from Pseudomonas aeruginosa . J Infect Dis 188:239–249
     [Google Scholar]
  9. Bröms J. E., Forslund A.-L., Forsberg Å., Francis M. S. 2003b; PcrH of Pseudomonas aeruginosa is essential for secretion and assembly of the type III translocon. J Infect Dis in press
    [Google Scholar]
  10. Cambronne E. D., Schneewind O. 2002; Yersinia enterocolitica type III secretion: yscM1 and yscM2 regulate yop gene expression by a posttranscriptional mechanism that targets the 5′ untranslated region of yop mRNA. J Bacteriol 184:5880–5893
     [Google Scholar]
  11. Cheng L. W., Schneewind O. 2000; Yersinia enterocolitica TyeA, an intracellular regulator of the type III machinery, is required for specific targeting of YopE, YopH, YopM, and YopN into the cytosol of eukaryotic cells. J Bacteriol 182:3183–3190
     [Google Scholar]
  12. Cornelis G. R., Boland A., Boyd A. P., Geuijen C., Iriarte M., Neyt C., Sory M. P., Stainier I. 1998; The virulence plasmid of Yersinia , an antihost genome. Microbiol Mol Biol Rev 62:1315–1352
     [Google Scholar]
  13. Dacheux D., Goure J., Chabert J., Usson Y., Attree I. 2001; Pore-forming activity of type III system-secreted proteins leads to oncosis of Pseudomonas aeruginosa -infected macrophages. Mol Microbiol 40:76–85
     [Google Scholar]
  14. Delahay R. M., Frankel G. 2002; Coiled-coil proteins associated with type III secretion systems: a versatile domain revisited. Mol Microbiol 45:905–916
     [Google Scholar]
  15. Dower W. J., Miller J. F., Ragsdale C. W. 1988; High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res 16:6127–6145
     [Google Scholar]
  16. Fällman M., Deleuil F., McGee K. 2002; Resistance to phagocytosis by Yersinia . Int J Med Microbiol 291:501–509
     [Google Scholar]
  17. 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]
  18. Francis M. S., Aili M., Wiklund M. L., Wolf-Watz H. 2000; A study of the YopD–LcrH interaction from Yersinia pseudotuberculosis reveals a role for hydrophobic residues within the amphipathic domain of YopD. Mol Microbiol 38:85–102
     [Google Scholar]
  19. Francis M. S., Lloyd S. A., Wolf-Watz H. 2001; The type III secretion chaperone LcrH co-operates with YopD to establish a negative, regulatory loop for control of Yop synthesis in Yersinia pseudotuberculosis . Mol Microbiol 42:1075–1093
     [Google Scholar]
  20. Frank D. W. 1997; The exoenzyme S regulon of Pseudomonas aeruginosa . Mol Microbiol 26:621–629
     [Google Scholar]
  21. Frithz-Lindsten E., Du Y., Rosqvist R., Forsberg Å. 1997; Intracellular targeting of exoenzyme S of Pseudomonas aeruginosa via type III-dependent translocation induces phagocytosis resistance, cytotoxicity and disruption of actin microfilaments. Mol Microbiol 25:1125–1139
     [Google Scholar]
  22. Frithz-Lindsten E., Holmström A., Jacobsson L., Soltani M., Olsson J., Rosqvist R., Forsberg Å. 1998; Functional conservation of the effector protein translocators PopB/YopB and PopD/YopD of Pseudomonas aeruginosa and Yersinia pseudotuberculosis . Mol Microbiol 29:1155–1165
     [Google Scholar]
  23. Fürste J. P., Pansegrau W., Frank R., Blöcker H., Scholz P., Bagdasarian M., Lanka E. 1986; Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene 48:119–131
     [Google Scholar]
  24. Håkansson S., Bergman T., Vanooteghem J. C., Cornelis G., Wolf-Watz H. 1993; YopB and YopD constitute a novel class of Yersinia Yop proteins. Infect Immun 61:71–80
     [Google Scholar]
  25. Håkansson S., Schesser K., Persson C., Galyov E. E., Rosqvist R., Homble F., Wolf-Watz H. 1996; The YopB protein of Yersinia pseudotuberculosis is essential for the translocation of Yop effector proteins across the target cell plasma membrane and displays a contact-dependent membrane disrupting activity. EMBO J 15:5812–5823
     [Google Scholar]
  26. Hanahan D. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
     [Google Scholar]
  27. Hartland E. L., Robins-Browne R. M. 1998; In vitro association between the virulence proteins, YopD and YopE, of Yersinia enterocolitica . FEMS Microbiol Lett 162:207–213
     [Google Scholar]
  28. Hartland E. L., Bordun A. M., Robins-Browne R. M. 1996; Contribution of YopB to virulence of Yersinia enterocolitica . Infect Immun 64:2308–2314
     [Google Scholar]
  29. Henriksson M. L., Francis M. S., Peden A., Aili M., Stefansson K., Palmer R., Aitken A., Hallberg B. 2002; A nonphosphorylated 14-3-3 binding motif on exoenzyme S that is functional in vivo . Eur J Biochem 269:4921–4929
     [Google Scholar]
  30. Hoiczyk E., Blobel G. 2001; Polymerization of a single protein of the pathogen Yersinia enterocolitica into needles punctures eukaryotic cells. Proc Natl Acad Sci U S A 98:4669–4674
     [Google Scholar]
  31. Holmström A., Pettersson J., Rosqvist R., Håkansson S., Tafazoli F., Fällman M., Magnusson K. E., Wolf-Watz H., Forsberg Å. 1997; YopK of Yersinia pseudotuberculosis controls translocation of Yop effectors across the eukaryotic cell membrane. Mol Microbiol 24:73–91
     [Google Scholar]
  32. Holmström A., Olsson J., Cherepanov P., Maier E., Nordfelth R., Pettersson J., Benz R., Wolf-Watz H., Forsberg Å. 2001; LcrV is a channel size-determining component of the Yop effector translocon of Yersinia . Mol Microbiol 39:620–632
     [Google Scholar]
  33. Horton R. M., Pease L. R. 1991; Recombination and mutagenesis of DNA sequences using PCR. In Directed Mutagenesis: a Practical Approach pp  217–247 Edited by McPherson M. J. New York: Oxford University Press;
     [Google Scholar]
  34. 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]
  35. Iriarte M., Sory M. P., Boland A., Boyd A. P., Mills S. D., Lambermont I., Cornelis G. R. 1998; TyeA, a protein involved in control of Yop release and in translocation of Yersinia Yop effectors. EMBO J 17:1907–1918
     [Google Scholar]
  36. Jin Q., He S. Y. 2001; Role of the Hrp pilus in type III protein secretion in Pseudomonas syringae . Science 294:2556–2558
     [Google Scholar]
  37. Kaniga K., Tucker S., Trollinger D., Galán J. E. 1995; Homologs of the Shigella IpaB and IpaC invasins are required for Salmonella typhimurium entry into cultured epithelial cells. J Bacteriol 177:3965–3971
     [Google Scholar]
  38. Kubori T., Matsushima Y., Nakamura D., Uralil J., Lara-Tejero M., Sukhan A., Galán J. E., Aizawa S. I. 1998; Supramolecular structure of the Salmonella typhimurium type III protein secretion system. Science 280:602–605
     [Google Scholar]
  39. Lyczak J. B., Cannon C. L., Pier G. B. 2000; Establishment of Pseudomonas aeruginosa infection: lessons from a versatile opportunist. Microbes Infect 2:1051–1060
     [Google Scholar]
  40. Matson J. S., Nilles M. L. 2002; Interaction of the Yersinia pestis type III regulatory proteins LcrG and LcrV occurs at a hydrophobic interface. BMC Microbiol 2, 16; Erratum 2:25
     [Google Scholar]
  41. Menard R., Sansonetti P., Parsot C., Vasselon T. 1994; Extracellular association and cytoplasmic partitioning of the IpaB and IpaC invasins of S. flexneri . Cell 79:515–525
     [Google Scholar]
  42. Milton D. L., O'Toole R., Horstedt P., Wolf-Watz H. 1996; Flagellin A is essential for the virulence of Vibrio anguillarum . J Bacteriol 178:1310–1319
     [Google Scholar]
  43. Neyt C., Cornelis G. R. 1999a; Insertion of a Yop translocation pore into the macrophage plasma membrane by Yersinia enterocolitica : requirement for translocators YopB and YopD, but not LcrG. Mol Microbiol 33:971–981
     [Google Scholar]
  44. Neyt C., Cornelis G. R. 1999b; Role of SycD, the chaperone of the Yersinia Yop translocators YopB and YopD. Mol Microbiol 31:143–156
     [Google Scholar]
  45. Nordfelth R., Wolf-Watz H. 2001; YopB of Yersinia enterocolitica is essential for YopE translocation. Infect Immun 69:3516–3518
     [Google Scholar]
  46. Pallen M. J., Dougan G., Frankel G. 1997; Coiled-coil domains in proteins secreted by type III secretion systems. Mol Microbiol 25:423–425
     [Google Scholar]
  47. Pettersson J., Nordfelth R., Dubinina E., Bergman T., Gustafsson M., Magnusson K. E., Wolf-Watz H. 1996; Modulation of virulence factor expression by pathogen target cell contact. Science 273:1231–1233
     [Google Scholar]
  48. Pettersson J., Holmström A., Hill J. 7 other authors 1999; The V-antigen of Yersinia is surface exposed before target cell contact and involved in virulence protein translocation. Mol Microbiol 32:961–976
     [Google Scholar]
  49. Price S. B., Cowan C., Perry R. D., Straley S. C. 1991; The Yersinia pestis V antigen is a regulatory protein necessary for Ca2(+)-dependent growth and maximal expression of low-Ca2+ response virulence genes. J Bacteriol 173:2649–2657
     [Google Scholar]
  50. Revell P. A., Miller V. L. 2001; Yersinia virulence: more than a plasmid. FEMS Microbiol Lett 205:159–164
     [Google Scholar]
  51. Rimpiläinen M., Forsberg Å., Wolf-Watz H. 1992; A novel protein, LcrQ, involved in the low-calcium response of Yersinia pseudotuberculosis shows extensive homology to YopH. J Bacteriol 174:3355–3363
     [Google Scholar]
  52. Rosqvist R., Forsberg Å., Wolf-Watz H. 1991; Intracellular targeting of the Yersinia YopE cytotoxin in mammalian cells induces actin microfilament disruption. Infect Immun 59:4562–4569
     [Google Scholar]
  53. Rosqvist R., Magnusson K. E., Wolf-Watz H. 1994; Target cell contact triggers expression and polarized transfer of Yersinia YopE cytotoxin into mammalian cells. EMBO J 13:964–972
     [Google Scholar]
  54. Rosqvist R., Håkansson S., Forsberg Å., Wolf-Watz H. 1995; Functional conservation of the secretion and translocation machinery for virulence proteins of yersiniae, salmonellae and shigellae. EMBO J 14:4187–4195
     [Google Scholar]
  55. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  56. Sarker M. R., Neyt C., Stainier I., Cornelis G. R. 1998; The Yersinia Yop virulon: LcrV is required for extrusion of the translocators YopB and YopD. J Bacteriol 180:1207–1214
     [Google Scholar]
  57. Sawa T., Yahr T. L., Ohara M., Kurahashi K., Gropper M. A., Wiener-Kronish J. P., Frank D. W. 1999; Active and passive immunization with the Pseudomonas V antigen protects against type III intoxication and lung injury. Nat Med 5:392–398
     [Google Scholar]
  58. Sekiya K., Ohishi M., Ogino T., Tamano K., Sasakawa C., Abe A. 2001; Supermolecular structure of the enteropathogenic Escherichia coli type III secretion system and its direct interaction with the EspA-sheath-like structure. Proc Natl Acad Sci U S A 98:11638–11643
     [Google Scholar]
  59. Simon R., Priefer U., Pühler A. 1983; A broad host range mobilisation system for in vivo genetic engineering: transposon mutagenesis in Gram negative bacteria. Biotechnology 1:787–796
     [Google Scholar]
  60. Skrzypek E., Straley S. C. 1995; Differential effects of deletions in lcrV on secretion of V antigen, regulation of the low-Ca2+ response, and virulence of Yersinia pestis . J Bacteriol 177:2530–2542
     [Google Scholar]
  61. Stover C. K., Pham X. Q., Erwin A. L. 28 other authors 2000; Complete genome sequence of Pseudomonas aeruginosa PA01, an opportunistic pathogen. Nature 406:959–964
     [Google Scholar]
  62. Straley S. C., Plano G. V., Skrzypek E., Haddix P. L., Fields K. A. 1993; Regulation by Ca2+ in the Yersinia low-Ca2+ response. Mol Microbiol 8:1005–1010
     [Google Scholar]
  63. Sundin C., Henriksson M. L., Hallberg B., Forsberg Å., Frithz-Lindsten E. 2001; Exoenzyme T of Pseudomonas aeruginosa elicits cytotoxicity without interfering with Ras signal transduction. Cell Microbiol 3:237–246
     [Google Scholar]
  64. Sundin C., Wolfgang M. C., Lory S., Forsberg Å., Frithz-Lindsten E. 2002; Type IV pili are not specifically required for contact dependent translocation of exoenzymes by Pseudomonas aeruginosa . Microb Pathog 33:265–277
     [Google Scholar]
  65. Tamano K., Aizawa S., Katayama E., Nonaka T., Imajoh-Ohmi S., Kuwae A., Nagai S., Sasakawa C. 2000; Supramolecular structure of the Shigella type III secretion machinery: the needle part is changeable in length and essential for delivery of effectors. EMBO J 19:3876–3887
     [Google Scholar]
  66. Tardy F., Homble F., Neyt C., Wattiez R., Cornelis G. R., Ruysschaert J. M., Cabiaux V. 1999; Yersinia enterocolitica type III secretion–translocation system: channel formation by secreted Yops. EMBO J 18:6793–6799
     [Google Scholar]
  67. 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]
  68. Vallis A. J., Yahr T. L., Barbieri J. T., Frank D. W. 1999; Regulation of ExoS production and secretion by Pseudomonas aeruginosa in response to tissue culture conditions. Infect Immun 67:914–920
     [Google Scholar]
  69. Wachter C., Beinke C., Mattes M., Schmidt M. A. 1999; Insertion of EspD into epithelial target cell membranes by infecting enteropathogenic Escherichia coli . Mol Microbiol 31:1695–1707
     [Google Scholar]
  70. Wattiau P., Bernier B., Deslee P., Michiels T., Cornelis G. R. 1994; Individual chaperones required for Yop secretion by Yersinia . Proc Natl Acad Sci U S A 91:10493–10497
     [Google Scholar]
  71. Williams A. W., Straley S. C. 1998; YopD of Yersinia pestis plays a role in negative regulation of the low-calcium response in addition to its role in translocation of Yops. J Bacteriol 180:350–358
     [Google Scholar]
  72. Yahr T. L., Mende-Mueller L. M., Friese M. B., Frank D. W. 1997; Identification of type III secreted products of the Pseudomonas aeruginosa exoenzyme S regulon. J Bacteriol 179:7165–7168
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26322-0
Loading
Dissection of homologous translocon operons reveals a distinct role for YopD in type III secretion by Yersinia pseudotuberculosis
Microbiology 149, 2615 (2003); https://doi.org/10.1099/mic.0.26322-0
/content/journal/micro/10.1099/mic.0.26322-0
/content/journal/micro/10.1099/mic.0.26322-0
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