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ExoT Induces Atypical Anoikis Apoptosis in Target Host Cells by Transforming Crk Adaptor Protein into a Cytotoxin


We have previously demonstrated that ExoT is both necessary and sufficient to induce potent apoptosis in host epithelial cells in a manner that depends primarily on its ADP-ribosyltransferase (ADPRT) domain activity. However, the molecular basis underlying ExoT/ADPRT-induced apoptosis remains unknown. In this study, we demonstrate that ExoT/ADPRT by targeting the adaptor protein Crk, transforms this innocuous cellular protein into a cytotoxin that induces atypical anoikis by disrupting the focal adhesion sites which in turn interferes with the integrin-mediated pro-survival signaling.


Vyšlo v časopise: ExoT Induces Atypical Anoikis Apoptosis in Target Host Cells by Transforming Crk Adaptor Protein into a Cytotoxin. PLoS Pathog 11(5): e32767. doi:10.1371/journal.ppat.1004934
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004934

Souhrn

We have previously demonstrated that ExoT is both necessary and sufficient to induce potent apoptosis in host epithelial cells in a manner that depends primarily on its ADP-ribosyltransferase (ADPRT) domain activity. However, the molecular basis underlying ExoT/ADPRT-induced apoptosis remains unknown. In this study, we demonstrate that ExoT/ADPRT by targeting the adaptor protein Crk, transforms this innocuous cellular protein into a cytotoxin that induces atypical anoikis by disrupting the focal adhesion sites which in turn interferes with the integrin-mediated pro-survival signaling.


Zdroje

1. Hauser AR, Cobb E, Bodi M, Mariscal D, Valles J, et al. (2002) Type III protein secretion is associated with poor clinical outcomes in patients with ventilator-associated pneumonia caused by Pseudomonas aeruginosa. Crit Care Med 30: 521–528. 11990909

2. Engel J, Balachandran P (2009) Role of Pseudomonas aeruginosa type III effectors in disease. Curr Opin Microbiol 12: 61–66. doi: 10.1016/j.mib.2008.12.007 19168385

3. Engel JN (2003) Molecular pathogenesis of acute Pseudomonas aeruginosa infections. In: Hauser A, Rello J, editors. Severe Infections Caused by Pseudomonas aeruginosa. New York City: Kluwer Academic/Plenum Press. pp. 201–230.

4. Salyers AA, Whitt DD, editors (1994) Bacterial pathogenesis: a molecular approach. Washington, D. C.: ASM Press. 260–268 p.

5. Stavrinides J, McCann HC, Guttman DS (2008) Host-pathogen interplay and the evolution of bacterial effectors. Cell Microbiol 10: 285–292. 18034865

6. Hueck C (1998) Type III protein secretion systems in bacterial pathogens of animals and plants. Microbiol and Molec Biol Rev 62: 379–433. 9618447

7. Hauser AR (2009) The type III secretion system of Pseudomonas aeruginosa: infection by injection. Nat Rev Microbiol 7: 654–665. doi: 10.1038/nrmicro2199 19680249

8. Sato H, Frank DW (2004) ExoU is a potent intracellular phospholipase. Mol Microbiol 53: 1279–1290. 15387809

9. Sato H, Feix JB, Hillard CJ, Frank DW (2005) Characterization of phospholipase activity of the Pseudomonas aeruginosa type III cytotoxin, ExoU. J Bacteriol 187: 1192–1195. 15659695

10. Henriksson ML, Sundin C, Jansson AL, Forsberg A, Palmer RH, et al. (2002) Exoenzyme S shows selective ADP-ribosylation and GTPase-activating protein (GAP) activities towards small GTPases in vivo. Biochem J 367: 617–628. 12132999

11. Shaver CM, Hauser AR (2004) Relative contributions of Pseudomonas aeruginosa ExoU, ExoS, and ExoT to virulence in the lung. Infect Immun 72: 6969–6977. 15557619

12. Garrity-Ryan L, Kazmierczak B, Kowal R, Commolli J, Hauser A, et al. (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. 11083836

13. Krall R, Schmidt G, Aktories K, Barbieri JT (2000) Pseudomonas aeruginosa ExoT is a Rho GTPase-activating protein. Infect Immun 68: 6066–6068. 10992524

14. Wurtele M, Wolf E, Pederson KJ, Buchwald G, Ahmadian MR, et al. (2001) How the Pseudomonas aeruginosa ExoS toxin downregulates Rac. Nat Struct Biol 8: 23–26. 11135665

15. Braun M, Stuber K, Schlatter Y, Wahli T, Kuhnert P, et al. (2002) Characterization of an ADP-ribosyltransferase toxin (AexT) from Aeromonas salmonicida subsp. salmonicida. J Bacteriol 184: 1851–1858. 11889090

16. Barbieri JT, Sun J (2004) Pseudomonas aeruginosa ExoS and ExoT. Rev Physiol Biochem Pharmacol 152: 79–92. 15375697

17. Sun J, Barbieri JT (2003) Pseudomonas aeruginosa ExoT ADP-ribosylates CT10 regulator of kinase (Crk) proteins. J Biol Chem 278: 32794–32800. 12807879

18. Garrity-Ryan L, Shafikhani S, Balachandran P, Nguyen L, Oza J, et al. (2004) The ADP ribosyltransferase domain of Pseudomonas aeruginosa ExoT contributes to its biological activities. Infection and immunity 72: 546–558. 14688136

19. Sato H, Frank DW, Hillard CJ, Feix JB, Pankhaniya RR, et al. (2003) The mechanism of action of the Pseudomonas aeruginosa-encoded type III cytotoxin, ExoU. Embo J 22: 2959–2969. 12805211

20. Barbieri JT (2000) Pseudomonas aeruginosa exoenzyme S, a bifunctional type-III secreted cytotoxin. Int J Med Microbiol 290: 381–387. 11111915

21. Yahr T, Mende-Mueller LM, Friese MB, Frank DW (1997) Identification of type III secreted products of the Pseudomonas aeruginosa exoenzyme S regulon. J Bacteriol 179: 7165–7168. 9371466

22. Vance RE, Rietsch A, Mekalanos JJ (2005) Role of the type III secreted exoenzymes S, T, and Y in systemic spread of Pseudomonas aeruginosa PAO1 in vivo. Infect Immun 73: 1706–1713. 15731071

23. 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: 13899–13904. 9811898

24. Feltman H, Khan S, Jain M, Peterson L, Hauser A (2001) Type III secretion genotypes of clinical and environmental Pseudmonas aeruginosa isolates. ASM abstracts: D22.

25. Feltman H, Schulert G, Khan S, Jain M, Peterson L, et al. (2001) Prevalence of type III secretion genes in clinical and environmental isolates of Pseudomonas aeruginosa. Microbiology 147: 2659–2669. 11577145

26. Balachandran P, Dragone L, Garrity-Ryan L, Lemus A, Weiss A, et al. (2007) The ubiquitin ligase Cbl-b limits Pseudomonas aeruginosa exotoxin T-mediated virulence. J Clin Invest 117: 419–427. 17235393

27. Shafikhani SH, Engel J (2006) Pseudomonas aeruginosa type III-secreted toxin ExoT inhibits host-cell division by targeting cytokinesis at multiple steps. Proceedings of the National Academy of Sciences of the United States of America 103: 15605–15610. 17030800

28. Shafikhani SH, Morales C, Engel J (2008) The Pseudomonas aeruginosa type III secreted toxin ExoT is necessary and sufficient to induce apoptosis in epithelial cells. Cellular microbiology 10: 994–1007. 18053004

29. Wood S, Pithadia R, Rehman T, Zhang L, Plichta J, et al. (2013) Chronic alcohol exposure renders epithelial cells vulnerable to bacterial infection. PLoS One 8: e54646. doi: 10.1371/journal.pone.0054646 23358457

30. Chiarugi P, Giannoni E (2008) Anoikis: a necessary death program for anchorage-dependent cells. Biochem Pharmacol 76: 1352–1364. doi: 10.1016/j.bcp.2008.07.023 18708031

31. Bouchard V, Harnois C, Demers MJ, Thibodeau S, Laquerre V, et al. (2008) B1 integrin/Fak/Src signaling in intestinal epithelial crypt cell survival: integration of complex regulatory mechanisms. Apoptosis 13: 531–542. doi: 10.1007/s10495-008-0192-y 18322799

32. Vachon PH (2011) Integrin signaling, cell survival, and anoikis: distinctions, differences, and differentiation. Journal of signal transduction 2011: 738137. doi: 10.1155/2011/738137 21785723

33. Cardone MH, Salvesen GS, Widmann C, Johnson G, Frisch SM (1997) The regulation of anoikis: MEKK-1 activation requires cleavage by caspases. Cell 90: 315–323. 9244305

34. Orford K, Orford CC, Byers SW (1999) Exogenous expression of beta-catenin regulates contact inhibition, anchorage-independent growth, anoikis, and radiation-induced cell cycle arrest. J Cell Biol 146: 855–868. 10459019

35. Fang D, Hawke D, Zheng Y, Xia Y, Meisenhelder J, et al. (2007) Phosphorylation of beta-catenin by AKT promotes beta-catenin transcriptional activity. J Biol Chem 282: 11221–11229. 17287208

36. Frisch SM, Schaller M, Cieply B (2013) Mechanisms that link the oncogenic epithelial-mesenchymal transition to suppression of anoikis. J Cell Sci 126: 21–29. doi: 10.1242/jcs.120907 23516327

37. Cheng TL, Lai CH, Jiang SJ, Hung JH, Liu SK, et al. (2014) RHBDL2 Is a Critical Membrane Protease for Anoikis Resistance in Human Malignant Epithelial Cells. Scientific World Journal 2014: 902987. doi: 10.1155/2014/902987 24977233

38. Puck TT, Marcus PI (1955) A Rapid Method for Viable Cell Titration and Clone Production with Hela Cells in Tissue Culture: The Use of X-Irradiated Cells to Supply Conditioning Factors. Proc Natl Acad Sci U S A 41: 432–437. 16589695

39. Goldufsky J, Wood S, Hajihossainlou B, Rehman T, Majdobeh O, et al. (2015) Pseudomonas aeruginosa exotoxin T induces potent cytotoxicity against a variety of murine and human cancer cell lines. J Med Microbiol 64: 164–173. doi: 10.1099/jmm.0.000003-0 25627204

40. Deng Q, Sun J, Barbieri JT (2005) Uncoupling Crk signal transduction by Pseudomonas exoenzyme T. J Biol Chem 280: 35953–35960. 16123042

41. Kar B, Reichman CT, Singh S, O'Connor JP, Birge RB (2007) Proapoptotic function of the nuclear Crk II adaptor protein. Biochemistry 46: 10828–10840. 17764157

42. Austgen K, Johnson ET, Park TJ, Curran T, Oakes SA (2012) The adaptor protein CRK is a pro-apoptotic transducer of endoplasmic reticulum stress. Nat Cell Biol 14: 87–92. doi: 10.1038/ncb2395 22179045

43. Smith JJ, Richardson DA, Kopf J, Yoshida M, Hollingsworth RE, et al. (2002) Apoptotic regulation by the Crk adapter protein mediated by interactions with Wee1 and Crm1/exportin. Mol Cell Biol 22: 1412–1423. 11839808

44. Parrizas M, Blakesley VA, Beitner-Johnson D, Le Roith D (1997) The proto-oncogene Crk-II enhances apoptosis by a Ras-dependent, Raf-1/MAP kinase-independent pathway. Biochem Biophys Res Commun 234: 616–620. 9175762

45. Evans EK, Lu W, Strum SL, Mayer BJ, Kornbluth S (1997) Crk is required for apoptosis in Xenopus egg extracts. Embo J 16: 230–241. 9029144

46. Cho SY, Klemke RL (2000) Extracellular-regulated kinase activation and CAS/Crk coupling regulate cell migration and suppress apoptosis during invasion of the extracellular matrix. J Cell Biol 149: 223–236. 10747099

47. Salameh A, Galvagni F, Bardelli M, Bussolino F, Oliviero S (2005) Direct recruitment of CRK and GRB2 to VEGFR-3 induces proliferation, migration, and survival of endothelial cells through the activation of ERK, AKT, and JNK pathways. Blood 106: 3423–3431. 16076871

48. Lamorte L, Royal I, Naujokas M, Park M (2002) Crk adapter proteins promote an epithelial-mesenchymal-like transition and are required for HGF-mediated cell spreading and breakdown of epithelial adherens junctions. Mol Biol Cell 13: 1449–1461. 12006644

49. Rodrigues SP, Fathers KE, Chan G, Zuo D, Halwani F, et al. (2005) CrkI and CrkII function as key signaling integrators for migration and invasion of cancer cells. Molecular cancer research: MCR 3: 183–194. 15831672

50. Park TJ, Boyd K, Curran T (2006) Cardiovascular and craniofacial defects in Crk-null mice. Mol Cell Biol 26: 6272–6282. 16880535

51. Kim SH, Turnbull J, Guimond S (2011) Extracellular matrix and cell signalling: the dynamic cooperation of integrin, proteoglycan and growth factor receptor. J Endocrinol 209: 139–151. doi: 10.1530/JOE-10-0377 21307119

52. Cabodi S, Di Stefano P, Leal Mdel P, Tinnirello A, Bisaro B, et al. (2010) Integrins and signal transduction. Adv Exp Med Biol 674: 43–54. 20549939

53. Frisch SM, Vuori K, Ruoslahti E, Chan-Hui PY (1996) Control of adhesion-dependent cell survival by focal adhesion kinase. J Cell Biol 134: 793–799. 8707856

54. Kanchanawong P, Shtengel G, Pasapera AM, Ramko EB, Davidson MW, et al. (2010) Nanoscale architecture of integrin-based cell adhesions. Nature 468: 580–584. doi: 10.1038/nature09621 21107430

55. Zaidel-Bar R, Milo R, Kam Z, Geiger B (2007) A paxillin tyrosine phosphorylation switch regulates the assembly and form of cell-matrix adhesions. J Cell Sci 120: 137–148. 17164291

56. Feller SM (2001) Crk family adaptors-signalling complex formation and biological roles. Oncogene 20: 6348–6371. 11607838

57. Almeida EA, Ilic D, Han Q, Hauck CR, Jin F, et al. (2000) Matrix survival signaling: from fibronectin via focal adhesion kinase to c-Jun NH(2)-terminal kinase. J Cell Biol 149: 741–754. 10791986

58. Turner CE (2000) Paxillin interactions. J Cell Sci 113 Pt 23: 4139–4140. 11069756

59. Owens TW, Valentijn AJ, Upton JP, Keeble J, Zhang L, et al. (2009) Apoptosis commitment and activation of mitochondrial Bax during anoikis is regulated by p38MAPK. Cell Death Differ 16: 1551–1562. doi: 10.1038/cdd.2009.102 19662026

60. Wei L, Yang Y, Zhang X, Yu Q (2004) Cleavage of p130Cas in anoikis. J Cell Biochem 91: 325–335. 14743392

61. Wozniak MA, Modzelewska K, Kwong L, Keely PJ (2004) Focal adhesion regulation of cell behavior. Biochim Biophys Acta 1692: 103–119. 15246682

62. Parsons JT (2003) Focal adhesion kinase: the first ten years. J Cell Sci 116: 1409–1416. 12640026

63. Nieto-Pelegrin E, Meiler E, Martin-Villa JM, Benito-Leon M, Martinez-Quiles N (2014) Crk adaptors negatively regulate actin polymerization in pedestals formed by enteropathogenic Escherichia coli (EPEC) by binding to Tir effector. PLoS Pathog 10: e1004022. doi: 10.1371/journal.ppat.1004022 24675776

64. Lee WL, Cosio G, Ireton K, Grinstein S (2007) Role of CrkII in Fcgamma receptor-mediated phagocytosis. J Biol Chem 282: 11135–11143. 17308335

65. Martinez-Quiles N, Feuerbacher LA, Benito-Leon M, Hardwidge PR (2014) Contribution of Crk Adaptor Proteins to Host Cell and Bacteria Interactions. Biomed Res Int 2014: 372901. doi: 10.1155/2014/372901 25506591

66. Wood S, Sivaramakrishnan G, Engel J, Shafikhani SH (2011) Cell migration regulates the kinetics of cytokinesis. Cell Cycle 10: 648–654. 21293189

67. Kumar A, Zloza A, Moon RT, Watts J, Tenorio AR, et al. (2008) Active beta-catenin signaling is an inhibitory pathway for human immunodeficiency virus replication in peripheral blood mononuclear cells. J Virol 82: 2813–2820. doi: 10.1128/JVI.02498-07 18199649

68. Shafikhani SH, Mostov K, Engel J (2008) Focal adhesion components are essential for mammalian cell cytokinesis. Cell Cycle 7: 2868–2876. 18787414

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Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

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