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Pseudomonas aeruginosa ExoS and ExoT

  • J. T. BarbieriEmail author
  • J. Sun
Chapter
Part of the Reviews of Physiology, Biochemistry and Pharmacology book series (REVIEWS, volume 152)

Abstract

ExoS and ExoT are bi-functional type-III cytotoxins of Pseudomonas aeruginosa that share 76% primary amino acid homology and contain N-terminal RhoGAP domains and C-terminal ADP-ribosylation domains. The Rho GAP activities of ExoS and ExoT appear to be biochemically and biologically identical, targeting Rho, Rac, and Cdc42. Expression of the RhoGAP domain in mammalian cells results in the disruption of the actin cytoskeleton and interference of phagocytosis. Expression of the ADP-ribosyltransferase domain of ExoS elicits a cytotoxic phenotype in cultured cells, while expression of ExoT appears to interfere with host cell phagocytic activity. Recent studies showed that ExoS and ExoT ADP-ribosylate different substrates. While ExoS has poly-substrate specificity and can ADP-ribosylate numerous host proteins, ExoT ADP-ribosylates a more restricted subset of host proteins including the Crk proteins. Protein modeling predicts that electrostatic interactions contribute to the substrate specificity of the ADP-ribosyltransferase domains of ExoS and ExoT.

Keywords

Pseudomonas Aeruginosa Multidrug Transporter Focal Adhesion Complex Active Site Loop RhoGAP Domain 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

GAP

GTPase activating protein

ADP-r

Adenosine diphosphate ribose

MLD

Membrane localization domain

FAS

Factor activating exoenzyme S

Crk

CT-10 regulator of kinase

SBTI

Soybean trypsin inhibitor

GEF

Guanine nucleotide exchange factor

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References

  1. Black DS, Marie-Cardine A, Schraven B, Bliska JB (2000) The Yersinia tyrosine phosphatase YopH targets a novel adhesion-regulated signalling complex in macrophages” Cell Microbiol 2(5):401–414PubMedCrossRefGoogle Scholar
  2. Blocker A, Jouihri N, Larquet E, Gounon P, Ebel F, Parsot C, Sansonetti P, Allaoui A (2001) Structure and composition of the Shigella flexneri “needle complex,” a part of its type III secreton” Mol Microbiol 39(3):652–663PubMedCrossRefGoogle Scholar
  3. Burton EA, Plattner R, Pendergast AM (2003) Abl tyrosine kinases are required for infection by Shigella flexneri” EMBO J 22(20):5471–5479PubMedCrossRefGoogle Scholar
  4. Coburn J (1992) Pseudomonas aeruginosa exoenzyme S. Curr Top Microbiol Immunol 175:133–143PubMedGoogle Scholar
  5. Coburn J and Gill DM (1991) ADP-ribosylation of p21ras and related proteins by Pseudomonas aeruginosa exoenzyme S. Infect Immun 59(11):4259–4262PubMedGoogle Scholar
  6. Coburn J, Dillon ST, Iglewski BH, Gill DM (1989) Exoenzyme S of Pseudomonas aeruginosa ADP-ribosylates the intermediate filament protein vimentin. Infect Immun 57(3):996–998PubMedGoogle Scholar
  7. Coburn J, Wyatt RT, Iglewski BH, Gill DM (1989) Several GTP-binding proteins, including p21c-H-ras, are preferred substrates of Pseudomonas aeruginosa exoenzyme S. J Biol Chem 264(15):9004–9008PubMedGoogle Scholar
  8. Coburn J, Kane AV, Feig L, Gill DM (1991) Pseudomonas aeruginosa exoenzyme S requires a eukaryotic protein for ADP-ribosyltransferase activity. J Biol Chem 266(10):6438–6446PubMedGoogle Scholar
  9. Cowell BA, Chen DY, Frank DW, Vallis AJ, Fleiszig SM (2000) ExoT of cytotoxic Pseudomonas aeruginosa prevents uptake by corneal epithelial cells” Infect Immun 68(1):403–406PubMedCrossRefGoogle Scholar
  10. Finck-Barbancon V, Goranson J, Zhu L, Sawa T, Wiener-Kronish JP, Fleiszig SM, Wu C, Mende-Mueller L, Frank DW (1997) ExoU expression by Pseudomonas aeruginosa correlates with acute cytotoxicity and epithelial injury. Mol Microbiol 25(3):547–557PubMedCrossRefGoogle Scholar
  11. Fleiszig SM, Wiener-Kronish JP, Miyazaki H, Vallas V, Mostov KE, Kanada D, Sawa T, Yen TS, Frank DW (1997) Pseudomonas aeruginosa-mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S. Infect Immun 65(2):579–586PubMedGoogle Scholar
  12. Frank DW (1997) The exoenzyme S regulon of Pseudomonas aeruginosa. Mol Microbiol 26(4):621–629PubMedCrossRefGoogle Scholar
  13. Frithz-Lindsten E, Du Y, Rosqvist R, Forsberg A (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(6):1125–1139PubMedCrossRefGoogle Scholar
  14. Fu H, Coburn J, Collier RJ (1993) The eukaryotic host factor that activates exoenzyme S of Pseudomonas aeruginosa is a member of the 14-3-3 protein family. Proc Natl Acad Sci U S A 90(6):2320–2324PubMedCrossRefGoogle Scholar
  15. Ganesan AK, Frank DW, Misra RP, Schmidt G, Barbieri JT (1998) Pseudomonas aeruginosa exoenzyme S ADP-ribosylates Ras at multiple sites. J Biol Chem 273(13):7332–7337PubMedCrossRefGoogle Scholar
  16. Ganesan AK, Mende-Mueller L, Selzer J, Barbieri JT (1999) Pseudomonas aeruginosa exoenzyme S, a double ADP-ribosyltransferase, resembles vertebrate mono-ADP-ribosyltransferases. J Biol Chem 274(14):9503–9508PubMedCrossRefGoogle Scholar
  17. Garrity-Ryan L, Kazmierczak B, Kowal R, Comolli J, Hauser A, Engel JN (2000) The arginine finger domain of ExoT contributes to actin cytoskeleton disruption and inhibition of internalization of Pseudomonas aeruginosa by epithelial cells and macrophages. Infect Immun 68(12):7100–7113PubMedCrossRefGoogle Scholar
  18. Geiser TK, Kazmierczak BI, Garrity-Ryan LK, Matthay MA, Engel JN (2001) Pseudomonas aeruginosa ExoT inhibits in vitro lung epithelial wound repair. Cell Microbiol 3(4):223–236PubMedCrossRefGoogle Scholar
  19. Goehring UM, Schmidt G, Pederson KJ, Aktories K, Barbieri JT (1999) The N-terminal domain of Pseudomonas aeruginosa exoenzyme S is a GTPase-activating protein for Rho GTPases. J Biol Chem 274(51):36369–36372PubMedCrossRefGoogle Scholar
  20. Han S, Craig JA, Putnam CD, Carozzi NB, Tainer JA (1999) Evolution and mechanism from structures of an ADP-ribosylating toxin and NAD complex. Nat Struct Biol 6(10):932–936PubMedCrossRefGoogle Scholar
  21. Han S, Arvai AS, Clancy SB, Tainer JA (2001) Crystal structure and novel recognition motif of rho ADP-ribosylating C3 exoenzyme from Clostridium botulinum: structural insights for recognition specificity and catalysis. J Mol Biol 305(1):95–107PubMedCrossRefGoogle Scholar
  22. Henriksson ML, Troller U, Hallberg B (2000) 14-3-3 proteins are required for the inhibition of Ras by exoenzyme S. Biochem J 349 Pt 3:697–701PubMedGoogle Scholar
  23. Iglewski BH, Sadoff J, Bjorn MJ, Maxwell ES (1978) Pseudomonas aeruginosa exoenzyme S: an adenosine diphosphate ribosyltransferase distinct from toxin A. Proc Natl Acad Sci U S A 75(7):3211–3215PubMedCrossRefGoogle Scholar
  24. Jia J, Alaoui-El-Azher M, Chow M, Chambers TC, Baker H, Jin S (2003) c-Jun NH2-terminal kinase-mediated signaling is essential for Pseudomonas aeruginosa ExoS-induced apoptosis. Infect Immun 71(6):3361–3370PubMedCrossRefGoogle Scholar
  25. Kaufman MR, Jia J, Zeng L, Ha U, Chow M, Jin S (2000) Pseudomonas aeruginosa mediated apoptosis requires the ADP-ribosylating activity of exoS. Microbiology 146 (Pt 10):2531–2541PubMedGoogle Scholar
  26. Kazmierczak BI and Engel JN (2002) Pseudomonas aeruginosa ExoT acts in vivo as a GTPase-activating protein for RhoA, Rac1, and Cdc42. Infect Immun 70(4):2198–2205PubMedCrossRefGoogle Scholar
  27. Knight DA, Finck-Barbancon V, Kulich SM, Barbieri JT (1995) Functional domains of Pseudomonas aeruginosa exoenzyme S. Infect Immun 63(8):3182–3186PubMedGoogle Scholar
  28. Krall R, Schmidt G, Aktories K, Barbieri JT (2000) Pseudomonas aeruginosa ExoT is a Rho GTPase-activating protein. Infect Immun 68(10):6066–6068PubMedCrossRefGoogle Scholar
  29. Krall R, Sun J, Pederson KJ, Barbieri JT (2002) In vivo rho GTPase-activating protein activity of Pseudomonas aeruginosa cytotoxin ExoS. Infect Immun 70(1):360–367PubMedCrossRefGoogle Scholar
  30. Krall R, Zhang Y, Barbieri JT (2004) Intracellular membrane localization of pseudomonas ExoS and Yersinia YopE in mammalian cells. J Biol Chem 279(4):2747–2753PubMedCrossRefGoogle Scholar
  31. Kulich SM, Frank DW, Barbieri JT (1993) Purification and characterization of exoenzyme S from Pseudomonas aeruginosa 388. Infect Immun 61(1):307–313PubMedGoogle Scholar
  32. Kulich SM, Yahr TL, Mende-Mueller LM, Barbieri JT, Frank DW (1994) Cloning the structural gene for the 49-kDa form of exoenzyme S (exoS) from Pseudomonas aeruginosa strain 388. J Biol Chem 269(14):10431–10437PubMedGoogle Scholar
  33. Lanotte P, Watt S, Mereghetti L, Dartiguelongue N, Rastegar-Lari A, Goudeau A, Quentin R (2004) Genetic features of Pseudomonas aeruginosa isolates from cystic fibrosis patients compared with those of isolates from other origins. J Med Microbiol 53(1):73–81PubMedCrossRefGoogle Scholar
  34. Liu S, Kulich SM, Barbieri JT (1996) Identification of glutamic acid 381 as a candidate active site residue of Pseudomonas aeruginosa exoenzyme S. Biochemistry 35(8):2754–2758PubMedCrossRefGoogle Scholar
  35. Liu S, Yahr TL, Frank DW, Barbieri JT (1997) Biochemical relationships between the 53-kilodalton (Exo53) and 49-kilodalton (ExoS) forms of exoenzyme S of Pseudomonas aeruginosa. J Bacteriol 179(5):1609–1613PubMedGoogle Scholar
  36. Lomholt JA, Poulsen K, Kilian M (2001) Epidemic population structure of Pseudomonas aeruginosa: evidence for a clone that is pathogenic to the eye and that has a distinct combination of virulence factors. Infect Immun 69(10):6284–6295PubMedCrossRefGoogle Scholar
  37. Marchalonis JJ, Schluter SF, Bernstein RM, Hohman VS (1998) Antibodies of sharks: revolution and evolution. Immunol Rev 166:103–122PubMedCrossRefGoogle Scholar
  38. Neyfakh AA (2002) Mystery of multidrug transporters: the answer can be simple. Mol Microbiol 44(5):1123–1130PubMedCrossRefGoogle Scholar
  39. Nicas, TI and Iglewski BH (1984) Isolation and characterization of transposon-induced mutants of Pseudomonas aeruginosa deficient in production of exoenzyme S. Infect Immun 45(2):470–474PubMedGoogle Scholar
  40. Pederson KJ, Vallis AJ, Aktories K, Frank DW, Barbieri JT (1999) The amino-terminal domain of Pseudomonas aeruginosa ExoS disrupts actin filaments via small-molecular-weight GTP-binding proteins. Mol Microbiol 32(2):393–401PubMedCrossRefGoogle Scholar
  41. Pederson KJ, Pal S, Vallis AJ, Frank DW, Barbieri JT (2000) Intracellular localization and processing of Pseudomonas aeruginosa ExoS in eukaryotic cells. Mol Microbiol 37(2):287–299PubMedCrossRefGoogle Scholar
  42. Pederson KJ, Krall R, Riese MJ, Barbieri JT (2002) Intracellular localization modulates targeting of ExoS, a type III cytotoxin, to eukaryotic signalling proteins. Mol Microbiol 46(5):1381–1390PubMedCrossRefGoogle Scholar
  43. Persson C, Carballeira N, Wolf-Watz H, Fallman M (1997) The PTPase YopH inhibits uptake of Yersinia, tyrosine phosphorylation of p130Cas and FAK, and the associated accumulation of these proteins in peripheral focal adhesions. EMBO J 16(9):2307–2318CrossRefGoogle Scholar
  44. Petosa C, Masters SC, Bankston LA, Pohl J, Wang B, Fu H, Liddington RC (1998) 14-3-3zeta binds a phosphorylated Raf peptide and an unphosphorylated peptide via its conserved amphipathic groove. J Biol Chem 273(26):16305–16310PubMedCrossRefGoogle Scholar
  45. Radke J, Pederson KJ, Barbieri JT (1999) Pseudomonas aeruginosa exoenzyme S is a biglutamic acid ADP-ribosyltransferase. Infect Immun 67(3):1508–1510PubMedGoogle Scholar
  46. Riese MJ and Barbieri JT (2002) Membrane localization contributes to the in vivo ADP-ribosylation of Ras by Pseudomonas aeruginosa ExoS. Infect Immun 70(4):2230–2232PubMedCrossRefGoogle Scholar
  47. Riese MJ, Wittinghofer A, Barbieri JT (2001) ADP ribosylation of Arg41 of Rap by ExoS inhibits the ability of Rap to interact with its guanine nucleotide exchange factor, C3G. Biochemistry 40(11):3289–3294PubMedCrossRefGoogle Scholar
  48. Riese MJ, Goehring UM, Ehrmantraut ME, Moss J, Barbieri JT, Aktories K, Schmidt G (2002) Auto-ADP-ribosylation of Pseudomonas aeruginosa ExoS. J Biol Chem 277(14):12082–12088PubMedCrossRefGoogle Scholar
  49. Sato H, Frank DW, Hillard CJ, Feix JB, Pankhaniya RR, Moriyama K, Finck-Barbancon V, Buchaklian A, Lei M, et al. (2003) The mechanism of action of the Pseudomonas aeruginosa-encoded type III cytotoxin, ExoU. EMBO J 22(12):2959–2969PubMedCrossRefGoogle Scholar
  50. Sun J and Barbieri JT (2003) Pseudomonas aeruginosa ExoT ADP-ribosylates CT10 regulator of kinase (Crk) proteins. J Biol Chem 278(35):32794–32800PubMedCrossRefGoogle Scholar
  51. Sun J and Barbieri JT (2004) How bacterial ADP-ribosylating toxins recognize substrates. Nat Struct Mol Biol (in press)Google Scholar
  52. Sundin C, Henriksson ML, Hallberg B, Forsberg A, Frithz-Lindsten E (2001) Exoenzyme T of Pseudomonas aeruginosa elicits cytotoxicity without interfering with Ras signal transduction. Cell Microbiol 3(4):237–246PubMedCrossRefGoogle Scholar
  53. Vallis AJ, Finck-Barbancon V, Yahr TL, Frank DW (1999) Biological effects of Pseudomonas aeruginosa type III-secreted proteins on CHO cells. Infect Immun 67(4):2040–2044PubMedGoogle Scholar
  54. Van Delden C and Iglewski BH (1998) Cell-to-cell signaling and Pseudomonas aeruginosa infections. Emerg Infect Dis 4(4):551–560PubMedCrossRefGoogle Scholar
  55. Wurtele M, Renault L, Barbieri JT, Wittinghofer A, Wolf E (2001) Structure of the ExoS GTPase activating domain. FEBS Lett 491(1–2):26–29PubMedCrossRefGoogle Scholar
  56. Wurtele M, Wolf E, Pederson KJ, Buchwald G, Ahmadian MR, Barbieri JT, Wittinghofer A (2001) How the Pseudomonas aeruginosa ExoS toxin downregulates Rac. Nat Struct Biol 8(1):23–26PubMedCrossRefGoogle Scholar
  57. Yahr TL, Barbieri JT, Frank DW (1996) Genetic relationship between the 53-and 49-kilodalton forms of exoenzyme S from Pseudomonas aeruginosa. J Bacteriol 178(5):1412–1419PubMedGoogle Scholar
  58. Yahr TL, Vallis AJ, Hancock MK, Barbieri JT, Frank DW (1998) ExoY, an adenylate cyclase secreted by the Pseudomonas aeruginosa type III system. Proc Natl Acad Sci U S A 95(23):13899–13904PubMedCrossRefGoogle Scholar
  59. Zhang L, Wang H, Liu D, Liddington R, Fu H (1997) Raf-1 kinase and exoenzyme S interact with 14-3-3zeta through a common site involving lysine 49. J Biol Chem 272(21):13717–13724PubMedCrossRefGoogle Scholar
  60. Zhang L, Wang H, Masters SC, Wang B, Barbieri JT, Fu H (1999) Residues of 14-3-3 zeta required for activation of exoenzyme S of Pseudomonas aeruginosa. Biochemistry 38(37):12159–12164PubMedCrossRefGoogle Scholar
  61. Zheleznova EE, Markham PN, Neyfakh AA, Brennan RG (1999) Structural basis of multidrug recognition by BmrR, a transcription activator of a multidrug transporter. Cell 96(3):353–362PubMedCrossRefGoogle Scholar
  62. Zheleznova EE, Markham P, Edgar R, Bibi E, Neyfakh AA, Brennan RG (2000) A structure-based mechanism for drug binding by multidrug transporters. Trends Biochem Sci 25(2):39–43PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Microbiology and Molecular GeneticsMedical College of WisconsinMilwaukeeUSA

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