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The Phosphoinositol-3-Kinase–Protein Kinase B/Akt Pathway Is Critical for Pseudomonas aeruginosa Strain PAK Internalization

Kierbel, A., Gassama-Diagne, A., Mostov, K., Engel, J. N.·2005·Molecular Biology of the Cell
DOI 10.1091/mbc.e04-08-0717
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Abstract

Several Pseudomonas aeruginosa strains are internalized by epithelial cells in vitro and in vivo, but the host pathways usurped by the bacteria to enter nonphagocytic cells are not clearly understood. Here, we report that internalization of strain PAK into epithelial cells triggers and requires activation of phosphatidylinositol 3-kinase (PI3K) and protein kinase B/Akt (Akt). Incubation of Madin-Darby canine kidney (MDCK) or HeLa cells with the PI3K inhibitors LY294002 (LY) or wortmannin abrogated PAK uptake. Addition of the PI3K product phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P 3 ] to polarized MDCK cells was sufficient to increase PAK internalization. PtdIns(3,4,5)P 3 accumulated at the site of bacterial binding in an LY-dependent manner. Akt phosphorylation correlated with PAK invasion. The specific Akt phosphorylation inhibitor SH-5 inhibited PAK uptake; internalization also was inhibited by small interfering RNA-mediated depletion of Akt phosphorylation. Expression of constitutively active Akt was sufficient to restore invasion when PI3K signaling was inhibited. Together, these results demonstrate that the PI3K signaling pathway is necessary and sufficient for the P. aeruginosa entry and provide the first example of a bacterium that requires Akt for uptake into epithelial cells.

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  1. Allewelt, M., Coleman, F. T., Grout, M., Priebe, G. P., and Pier, G. B. (2000). Acquisition of expression of the Pseudomonas aeruginosa ExoU cytotoxin leads to increased bacterial virulence in a murine model of acute pneumonia and systemic spread. Infect. Immun. 68 , 3998-4004. · DOI · Paper page
  2. Barbieri, A. M., Sha, Q., Bette-Bobillo, P., Stahl, P. D., and Vidal, M. (2001). ADP-ribosylation of Rab5 by ExoS of Pseudomonas aeruginosa affects endocytosis. Infect. Immun. 69 , 5329-5334. · DOI
  3. Bette-Bobillo, P., Giro, P., Sainte-Marie, J., and Vidal, M. (1998). Exoenzyme S from P. aeruginosa ADP ribosylates rab4 and inhibits transferrin recycling in SLO-permeabilized reticulocytes. Biochem. Biophys. Res. Commun. 244 , 336-341. · doi:10.1006/bbrc.1998.8263
  4. Burgering, B. M., and Coffer, P. J. (1995). Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature 376 , 599-602. · DOI · Paper page
  5. Celli, J., Olivier, M., and Finlay, B. B. (2001). Enteropathogenic Escherichia coli mediates antiphagocytosis through the inhibition of PI 3-kinase-dependent pathways. EMBO J. 20 , 1245-1258. · DOI
  6. Chi, E., Mehl, T., Nunn, D., and Lory, S. (1991). Interaction of Pseudomonas aeruginosa with A549 pneumocyte cells. Infect. Immun. 59 , 822-828. · DOI · Paper page
  7. Coffer, P. J., Jin, J., and Woodgett, J. R. (1998). Protein kinase B (c-Akt): a multifunctional mediator of phosphatidylinositol 3-kinase activation. Biochem J. 335 , 1-13. · DOI · Paper page
  8. Coombes, B. K., and Mahony, J. B. (2002). Identification of MEK- and phosphoinositide 3-kinase-dependent signalling as essential events during Chlamydia pneumoniae invasion of HEp2 cells. Cell Microbiol. 4 , 447-460. · DOI
  9. Cox, D., Tseng, C. C., Bjekic, G., and Greenberg, S. (1999). A requirement for phosphatidylinositol 3-kinase in pseudopod extension. J. Biol. Chem. 274 , 1240-1247. · DOI · Paper page
  10. Dacheux, D., Goure, J., Chabert, J., Usson, Y., and Attree, I. (2001). Poreforming activity of type III system-secreted proteins leads to oncosis of Pseudomonas aeruginosa -infected macrophages. Mol. Microbiol. 40 , 76-85. · DOI
  11. de Bentzmann, S., Roger, P., Dupuit, F., Bajolet-Laudinat, O., Fuchey, C., Plotkowski, M. C., and Puchelle, E. (1996). Asialo GM1 is a receptor for Pseudomonas aeruginosa adherence to regenerating respiratory epithelial cells. Infect. Immun. 64 , 1582-1588. · doi:10.1128/iai.64.5.1582-1588.1996
  12. de Bentzmann, S., Roger, P., and Puchelle, E. (1996). Pseudomonas aeruginosa adherence to remodelling respiratory epithelium. Eur. Resp. J. 10 , 2145-2150. · doi:10.1183/09031936.96.09102145
  13. Engel, J. N. (2003). Molecular Pathogenesis of acute Pseudomonas aeruginosa infections. In: Severe Infections Caused by Pseudomonas aeruginosa , ed. A. Hauser and J. Rello, New York: Kluwer Academic/Plenum Press, 201-230. · DOI
  14. Esen, M., Grassme, H., Riethmuller, J., Riehle, A., Fassbender, K., and Gulbins, E. (2001). Invasion of human epithelial cells by Pseudomonas aeruginosa involves src-like tyrosine kinases p60Src and p59Fyn. Infect. Immun. 69 , 281-287. · DOI
  15. Evans, D. J., Frank, D. W., Finck-Barbançon, V., Wu, C., and Fleiszig, S. M. (1998). Pseudomonas aeruginosa invasion and cytotoxicity are independent events, both of which involve protein tyrosine kinase activity. Infect. Immun. 66 , 1453-1459. · DOI
  16. Fleiszig, S. M., Evans, D. J., Do, N., Vallas, V., Shin, S., and Mostov, K. (1997a). Epithelial cell polarity affects susceptibility to Pseudomonas aeruginosa invasion and cytotoxicity. Infect. Immun. 65 , 2861-2867. · DOI · Paper page
  17. Fleiszig, S. M., Vallas, V., Jun, C., Mok, L., Balkovetz, D., Roth, M., and Mostov, K. (1998). Susceptibility of epithelial cells to Pseudomonas aeruginosa invasion and cytotoxicity is upregulated by hepatocyte growth factor. Infect. Immun. 66 , 3443-3446. · doi:10.1128/iai.66.7.3443-3446.1998
  18. Fleiszig, S. M., Zaidi, T. S., Fletcher, E. L., Preston, M. J., and Pier, G. B. (1994). Pseudomonas aeruginosa invades corneal epithelial cells during experimental infection. Infect. Immun. 62 , 3485-3493. · doi:10.1128/iai.62.8.3485-3493.1994
  19. Fleiszig, S. M., Zaidi, T. S., and Pier, G. B. (1995). Pseudomonas aeruginosa invasion of and multiplication within corneal epithelial cells in vitro. Infect. Immun. 63 , 4072-4077. · doi:10.1128/iai.63.10.4072-4077.1995
  20. Fleiszig, S.M.J., Wiener-Kronish, J. P., Miyazaki, H., Vallas, V., Mostov, K., Kanada, D., Sawa, T., Yen, T.S.B., and Frank, D. (1997b). Pseudomonas aeruginosa -mediated cytotoxicity and invasion correlate with distinct genotypes at the loci encoding exoenzyme S. Infect. Immun. 65 , 579-586. · DOI · Paper page
  21. Fraylick, J. E., Riese, M. J., Vincent, T. S., Barbieri, J. T., and Olson, J. C. (2002). ADP-ribosylation and functional effects of Pseudomonas exoenzyme S on cellular RalA. Biochemistry 41 , 9680-9687. · DOI
  22. Ganesan, A. K., Frank, D. W., Misra, R. P., Schmidt, G., and Barbieri, J. T. (1998). Pseudomonas aeruginosa exoenzyme S ADP-ribosylates Ras at multiple sites. J. Biol. Chem. 273 , 7332-7337. · DOI
  23. Ganesan, A. K., Vincent, T. S., Olson, J. C., and Barbieri, J. T. (1999). Pseudomonas aeruginosa exoenzyme S disrupts Ras-mediated signal transduction by inhibiting guanine nucleotide exchange factor-catalyzed nucleotide exchange. J. Biol. Chem. 274 , 21823-21829. · DOI
  24. Garrity-Ryan, L., Kazmierczak, B., Kowal, R., Commolli, J., Hauser, A., and Engel, J. (2000). The arginine finger domain of ExoT is required for actin cytoskeleton disruption and inhibition of internalization of Pseudomonas aeruginosa by epithelial cells and macrophages. Infect. Immun. 68 , 7100-7113. · DOI · Paper page
  25. Garrity-Ryan, L. et al . (2004). The ADP ribosyltransferase domain of Pseudomonas aeruginosa ExoT contributes to its biological activities. Infect. Immun. 72 , 546-558. · doi:10.1128/iai.72.1.546-558.2004
  26. Goehring, U. M., Schmidt, G., Pederson, K. J., Aktories, K., and Barbieri, J. T. (1999). The N-terminal domain of Pseudomonas aeruginosa exoenzyme S is a GTPase-activating protein for Rho GTPases. J. Biol. Chem. 274 , 36369-36372. · DOI · Paper page
  27. Grassmé, H., Kirschnek, S., Riethmueller, J., Andrea Riehle, von Kürthy, G., Lang, F., Weller, M., and GulbinS, E. (2000). CD95/CD95 ligand interactions on epithelial cells in host defense to Pseudomonas aeruginosa . Science 290 , 527-530. · DOI · Paper page
  28. Ha, U., and Jin, S. (2001). Growth phase-dependent invasion of Pseudomonas aeruginosa and its survival within HeLa cells. Infect. Immun. 69 , 4398-4406. · doi:10.1128/iai.69.7.4398-4406.2001
  29. Hauser, A. R., Kang, P. J., and Engel, J. (1998). PepA, a novel secreted protein of Pseudomonas aeruginosa , is necessary for cytotoxicity and virulence. Mol. Microbiol. 27 , 807-818. · DOI · Paper page
  30. Henriksson, M. L., Rosqvist, R., Telepnev, M., Wolf-Watz, H., and Hallberg, B. (2000). Ras effector pathway activation by EGF is inhibited in vivo by exoenzyme S ADP-ribosylation of Ras. Biochem. J. 347 , 217-222. · DOI
  31. Hirakata, Y. et al . (1998). Adherence to and penetration of human intestinal Caco-2 epithelial cell monolayers by Pseudomonas aeruginosa . Infect. Immun. 66 , 1748-1751. · doi:10.1128/iai.66.4.1748-1751.1998
  32. Ireton, K., Payrastre, B., Chap, H., Ogawa, W., Sakaue, H., Kasuga, M., and Cossart, P. (1996). A role for phosphoinositide 3-kinase in bacterial invasion. Science 274 , 780-782 [correction published in Science (1997) 275 , 464]. · DOI · Paper page
  33. Kazmierczak, B., and Engel, J. (2002). Pseudomonas aeruginosa ExoT acts in vivo as a GTPase activating protein for RhoA, Rac1, and Cdc42. Infect. Immun. 70 , 2198-2205. · DOI
  34. Kazmierczak, B. I., Jou, T.S., Mostov, K., and Engel, J. (2001). Rho-GTPase activity modulates Pseudomonas aeruginosa internalization by epithelial cells. Cell Microbiol. 3 , 85-98. · DOI
  35. 10.1091/mbc.e03-08-0559 · DOI
  36. Kozikowski, A. P., Sun, H., Brognard, J., and Dennis, P. A. (2003). Novel PI analogues selectively block activation of the pro-survival serine/threonine kinase Akt. J. Am. Chem. Soc. 125 , 1144-1145. · DOI · Paper page
  37. Krall, R., Schmidt, G., Aktories, K., and Barbieri, J. T. (2000). Pseudomonas aeruginosa ExoT is a Rho GTPase-activating protein. Infect. Immun. 68 , 6066-6068. · DOI · Paper page
  38. Kwok, T., Backert, S., Schwarz, H., Berger, J., and Meyer, T. F. (2002). Specific entry of Helicobacter pylori into cultured gastric epithelial cells via a zipper-like mechanism. Infect. Immun. 70 , 2108-2120. · DOI · Paper page
  39. Lemmon, M. A. (2003). Phosphoinositide recognition domains. Traffic 4 , 201-213. · DOI · Paper page
  40. Marshall, J. G., Booth, J. W., Stambolic, V., Mak, T., Balla, T., Schreiber, A. D., Meyer, T., and Grinstein, S. (2001). Restricted accumulation of phosphatidylinositol 3-kinase products in a plasmalemmal subdomain during Fc gamma receptor-mediated phagocytosis. J. Cell Biol. 153 , 1369-1380. · DOI · Paper page
  41. Martinez, J. J., and Hultgren, S. J. (2002). Requirement of Rho-family GTPases in the invasion of Type 1-piliated uropathogenic Escherichia coli . Cell Microbiol. 4 , 19-28. · DOI
  42. Martinez, J. J., Mulvey, M. A., Schilling, J. D., Pinkner, J. S., and Hultgren, S. J. (2000). Type 1 pilus-mediated bacterial invasion of bladder epithelial cells. EMBO J. 19 , 2803-2812. · DOI · Paper page
  43. Nobes, C. D., and Hall, A. (1995). Rho, Rac, and Cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell 81 , 53-62. · DOI · Paper page
  44. Ojcius, D. M., Bravo de Alba, Y., Kanellopoulos, J. M., Hawkins, R. A., Kelly, K. A., Rank, R. G., and Dautry-Varsat, A. (1998). Internalization of Chlamydia by dendritic cells and stimulation of Chlamydia -specific T cells. J. Immunol. 160 , 1297-1303. · DOI
  45. Ozaki, S., DeWald, D. B., Shope, J. C., Chen, J., and Prestwich, G. D. (2000). Intracellular delivery of phosphoinositides and inositol phosphates using polyamine carriers. Proc. Natl. Acad. Sci. USA 97 , 11286-11291. · DOI
  46. Pederson, K. J., Vallis, A. J., Aktories, K., Frank, D. W., and Barbieri, J. T. (1999). The amino-terminal domain of Pseudomonas aeruginosa ExoS disrupts actin filaments via small-molecular-weight GTP-binding proteins. Mol. Microbiol. 32 , 393-401. · DOI
  47. Pier, G. B., Grout, M., and Zaidi, T. S. (1997). Cystic fibrosis transmembrane conductance regulator is an epithelial cell receptor for clearance of Pseudomonas aeruginosa from the lung. Proc. Nat. Acad. Sci. USA 94 , 12088-12093. · DOI · Paper page
  48. Pizarro-Cerda, J., and Cossart, P. (2004). Subversion of phosphoinositide metabolism by intracellular bacterial pathogens. Nat. Cell Biol. 6 , 1026-1033. · DOI
  49. Reddy, M. A., Prasadarao, N. V., Wass, C. A., and Kim, K. S. (2000). Phosphatidylinositol 3-kinase activation and interaction with focal adhesion kinase in Escherichia coli K1 invasion of human brain microvascular endothelial cells. J. Biol. Chem. 275 , 36769-36774. · DOI
  50. Ren, X. D., and Schwartz, M. A. (1998). Regulation of inositol lipid kinases by Rho and Rac. Curr. Opin. Genet. Dev. 8 , 63-67. · DOI
  51. Riese, M. J., Wittinghofer, A., and Barbieri, J. T. (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 , 3289-3294. · doi:10.1021/bi002729q
  52. Roger, P., Puchelle, E., Bajolet-Laudinat, O., Tournier, J. M., Debordeaux, C., Plotkowski, M. C., Cohen, J. H., Sheppard, D., and de Bentzmann, S. (1999). Fibronectin and α5β1 integrin mediate binding of Pseudomonas aeruginosa to repairing airway epithelium. Eur. Respir. J. 13 , 1301-1309. · DOI
  53. Sato, H. et al . (2003). The mechanism of action of the Pseudomonas aeruginosa -encoded type III cytotoxin, ExoU. EMBO J. 22 , 2959-2969. · DOI · Paper page
  54. Servant, G., Weiner, O. D., Herzmark, P., Balla, T., Sedat, J. W., and Bourne, H. R. (2000). Polarization of chemoattractant receptor signaling during neutrophil chemotaxis. Science 287 , 1037-1040. · DOI · Paper page
  55. Steele-Mortimer, O., Knodler, L. A., Marcus, S. L., Scheid, M. P., Goh, B., Pfeifer, C. G., Duronio, V., and Finlay, B. B. (2000). Activation of Akt/protein kinase B in epithelial cells by the Salmonella typhimurium effector sigD. J. Biol. Chem. 275 , 37718-37724. · DOI · Paper page
  56. Stokoe, D., Stephens, L. R., Copeland, T., Gaffney, P. R., Reese, C. B., Painter, G. F., Holmes, A. B., McCormick, F., and Hawkins, P. T. (1997). Dual role of phosphatidylinositol-3,4,5-trisphosphate in the activation of protein kinase B. Science 277 , 567-570. · DOI · Paper page
  57. Sun, J., and Barbieri, J. T. (2003). Pseudomonas aeruginosa ExoT ADP-ribosylates CT10-regulator of kinase (Crk) proteins. J. Biol. Chem. 278 , 32794-32800. · DOI
  58. Tsang, K.W.T., Rutman, A., Tanaka, E., Lundt, V., Dewar, A., Cole, P. J., and Wilson, R. (1994). Interaction of Pseudomonas aeruginosa with human respiratory mucosa in vitro. Eur. Respir. J. 7 , 1746-1753. · DOI
  59. Ui, M., Okada, T., Hazeki, K., and Hazeki, O. (1995). Wortmannin as a unique probe for an intracellular signalling protein, phosphoinositide 3-kinase. Trends Biochem. Sci. 20 , 303-307. · DOI · Paper page
  60. Vanhaesebroeck, B., and Alessi, D. R. (2000). The PI3K-PDK1 connection: more than just a road to PKB. Biochem. J. 346 , 561-576. · doi:10.1042/bj3460561 · Paper page
  61. Vlahos, C. J., Matter, W. F., Hui, K. Y., and Brown, R. F. (1994). A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002). J. Biol. Chem. 269 , 5241-5248. · DOI · Paper page
  62. Watton, S. J., and Downward, J. (1999). Akt/PKB localisation and 3′ phosphoinositide generation at sites of epithelial cell-matrix and cell-cell interaction. Curr. Biol. 9 , 433-436. · DOI · Paper page
  63. Weiner, O. D., Neilsen, P. O., Prestwich, G. D., Kirschner, M. W., Cantley, L. C., and Bourne, H. R. (2002). A PtdInsP(3)- and Rho GTPase-mediated positive feedback loop regulates neutrophil polarity. Nat. Cell Biol. 4 , 509-513. · DOI · Paper page
  64. Wilkowsky, S. E., Barbieri, M. A., Stahl, P., and Isola, E. L. (2001). Trypanosoma cruzi: phosphatidylinositol 3-kinase and protein kinase B activation is associated with parasite invasion. Exp. Cell Res. 264 , 211-218. · doi:10.1006/excr.2000.5123
  65. Yamaguchi, T., and Yamada, H. (1991). Role of mechanical injury on airway surface in the pathogenesis of Pseudomonas aeruginosa . Am. Rev. Respir. Dis. 144 , 1147-1152. · DOI
  66. 10.1091/mbc.e02-06-0350 · DOI
  67. Zaas, D., Duncan, M., Li, G., Wright, J. R., and Abraham, S. N. (2005). Pseudomonas invasion of type I pneumocytes is dependent on the expression and phosphorylation of caveolin-2. J. Biol. Chem. 280 , 4864-4872. · DOI
  68. Zahm, J. M., Chevillard, M., and Puchelle, E. (1991). Wound repair of human surface respiratory epithelium. Am. J. Respir. Cell. Mol. Biol. 5 , 242-248. · doi:10.1165/ajrcmb/5.3.242
  69. Zenni, M. K., Giardina, P. C., Harvey, H. A., Shao, J., Ketterer, M. R., Lubaroff, D. M., Williams, R. D., and Apicella, M. A. (2000). Macropinocytosis as a mechanism of entry into primary human urethral epithelial cells by Neisseria gonorrhoeae . Infect. Immun. 68 , 1696-1699. · DOI

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Metadata

Authors
Kierbel, A., Gassama-Diagne, A., Mostov, K., & Engel, J. N.
Affiliations
  • Department of Medicine, University of California, San Francisco, San Francisco, CA 94143
  • Department of Anatomy, University of California, San Francisco, San Francisco, CA 94143
Venue
Molecular Biology of the Cell
Publisher
American Society for Cell Biology (ASCB)
Published
2005-05
Volume
16
Issue
5
Pages
2577-2585
ISSN
  • 1059-1524
  • 1939-4586
DOI
10.1091/mbc.e04-08-0717
Cited by
135
References
1
Language
en

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Kierbel, A., Gassama-Diagne, A., Mostov, K., & Engel, J. N. (2005). The Phosphoinositol-3-Kinase–Protein Kinase B/Akt Pathway Is Critical for Pseudomonas aeruginosa Strain PAK Internalization. Molecular Biology of the Cell. https://doi.org/10.1091/mbc.e04-08-0717

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