Antonie van Leeuwenhoek

, Volume 74, Issue 4, pp 191–197 | Cite as

Molecular bases of epithelial cell invasion by Shigella flexneri

  • Philippe J. Sansonetti
  • Coumaran Egile


The pathogenesis of shigellosis is characterized by the capacity of the causative microorganism, Shigella, to invade the epithelial cells that compose the mucosal surface of the colon in humans. The invasive process encompasses several steps which can be summarized as follows: entry of bacteria into epithelial cells involves signalling pathways that elicit a macropinocitic event. Upon contact with the cell surface, S. flexneri activates a Mxi/Spa secretory apparatus encoded by two operons comprising about 25 genes located on a large virulence plasmid of 220 kb. Through this specialized secretory apparatus, Ipa invasins are secreted, two of which (IpaB, 62 kDa and IpaC, 42 kDa) form a complex which is itself able to activate entry via its interaction with the host cell membrane. Interaction of this molecular complex with the cell surface elicits major rearrangements of the host cell cytoskeleton, essentially the polymerization of actin filaments that form bundles supporting the membrane projections which achieve bacterial entry. Active recruitment of the protooncogene pp 60c-src has been demonstrated at the entry site with consequent phosphorylation of cortactin. Also, the small GTPase Rho is controlling the cascade of signals that allows elongation of actin filaments from initial nucleation foci underneath the cell membrane. The regulatory signals involved as well as the proteins recruited indicate that Shigella induces the formation of an adherence plaque at the cell surface in order to achieve entry. Once intracellular, the bacterium lyses its phagocytic vacuole, escapes into the cytoplasm and starts moving the inducing polar, directed polymerization of actin on its surface, due to the expression of IcsA, a 120 kDa outer membrane protein, which is localized at one pole of the microorganism, following cleavage by SopA, a plasmid-encoded surface protease. In the context of polarized epithelial cells, bacteria then reach the intermediate junction and engage their components, particularly the cadherins, to form a protrusion which is actively internalized by the adjacent cell. Bacteria then lyse the two membranes, reach the cytoplasmic compartment again, and resume actin-driven movement.

epithelial cell invasion Shigella flexneri pathogenesis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Adam T, Arpin M, Pr & #x00E9;vost MC, Gounon P & Sansonetti PJ (1995) Cytoskeletal rearrangements and the functional role of T-plastin during entry of Shigella flexneri into HeLa cells. J. Cell. Biol. 129: 367–381Google Scholar
  2. Adam T, Giry M, Boquet P & Sansonetti PJ (1996) Rho-dependent membrane folding causes Shigella entry into epithelial cells. EMBO J. 15: 3315–3321Google Scholar
  3. Allaoui A, Mounier J, Pr & #x00E9;vost MC, Sansonetti PJ & Parsot C (1992) icsB: a Shigella flexneri virulence gene necessary for the lysis of protrusions during intercellular spread. Mol.Microbiol. 6: 1605–1616Google Scholar
  4. Bahrani FK, Sansonetti PJ & Parsot C (1997) Secretion of Ipa proteins by Shigella flexneri: inducing molecules and kinetics of activation. Infect. Immmun 65: 4005–4010Google Scholar
  5. Bernardini ML, Mounier J, d & #x2019;Hauteville H, Coquis-Rondon M & Sansonetti PJ (1989) Identification of icsA, a plasmid locus of Shigella flexneri which governs bacterial intra-and intercellular spread through interaction with F-actin. Proc. Natl. Acad. Sci. USA 86: 3867–3871Google Scholar
  6. Clerc P & Sansonetti PJ (1987) Entry of Shigella flexneri into HeLa cells: evidence for directed phagocytosis involving actin polymerization and myosin accumulation. Infect. Immun. 55: 2681–2688Google Scholar
  7. De Geyter C, Vogt B, Benjelloun-Touimi Z, Sansonetti PJ, Ruysschaert J-M, Parsot C & Cabiaux V (1997) Interaction of IpaC, a protein involved in entry of S. flexneri into epithelial cells, with lipid membranes. FEBS Lett. 400: 149–154Google Scholar
  8. Dehio C, Pr & #x00E9;vost MC & Sansonetti PJ (1995) Invasion of epithelial cells by Shigella flexneri induces tyrosine phosphorylation of cortactin by a pp60c?sr c mediated signalling pathway. EMBO J. 14: 2471–2482Google Scholar
  9. Domann E, Wehland J, Rohde M, Pistor S, Hartl M, Goebel M, Leimeister-Wachter M, Wuenscher M & Chakroborty T (1992) A novel bacterial virulence gene in Listeria monocytogenes required for host cell microfilament interaction with homology to the proline-rich region of vinculin. EMBO J. 11: 1981–1990Google Scholar
  10. & #x00C9;gile C, d & #x2019;Hauteville H, Parsot C & Sansonetti PJ (1997) SopA, an outer membrane protease achieving secretion and polar localization of IcsA in S. flexneri. Mol. Microbiol. 23: 1063–1073Google Scholar
  11. Francis CL, Ryan TA, Jones BD, Smith SJ & Falkows S (1993) Ruffles induced by Salmonella and other stimuli direct micropinocytosis of bacteria. Nature 364: 639–642Google Scholar
  12. Fukuda I, Suzuki T, Munakata H, Hayashi N, Katayama E, Yoshikawa M & Sasakawa C (1995) Cleavage of Shigella surface protein VirG occurs at a specific site, but the secretion is not essential for intracellular spreading. J. Bacteriol. 177: 1719–1726Google Scholar
  13. Garcia del Portillo F, Zwick MB, Leung KY & Finlay BB (1993) Salmonella induces the formation of filamentous structures containing lysosomal membrane glycoproteins in epithelial cells. Proc. Natl. Acad. Sci. USA 90: 10544–10548Google Scholar
  14. Goldberg MB (1997) Shigella actin-based motility in the absence of vinculin. Cell. Motil. Cytoskeleton 37: 44–53Google Scholar
  15. Goldberg MB & Th & #x00E9;riot JA (1995) Shigella flexneri surface protein IcsA is sufficient to direct actin-based motility. Proc. Natl. Acad. Sci. USA 92: 6572–6576Google Scholar
  16. Goldberg MB, Th & #x00E9;riot JA & Sansonetti PJ (1993) Regulation of surface presentation of IcsA, a Shigella protein essential to intracellular movement and spread, is growth phase dependent. Infect. Immun. 62: 5664–5668Google Scholar
  17. d & #x2019;Hauteville H & Sansonetti PJ (1992) Phosphorylation of IcsA by cAMP-dependent protein kinase and its effect on intercellular spread of Shigella flexneri. Mol. Microbiol. 6: 833–841Google Scholar
  18. High N, Mounier J, Pr & #x00E9;vost MC & Sansonetti PJ (1992) IpaB of Shigella flexneri causes entry into epithelial cells and escape from the phagocytic vacuole. EMBO J. 11: 1991–1999Google Scholar
  19. Isberg R (1991) Discrimination between intracellular uptake and surface adhesion of bacterial pathogens. Science 252: 934–938Google Scholar
  20. Kocks C, Gouin E, Tabouret M, Berche P, Ohayon H & Cossart P (1992) Listeria monocytogenes-induced actin assembly requires the actA gene product, a surface protein. Cell 68: 521–531Google Scholar
  21. Kocks C, Marchand JB, Gouin E, d & #x2019;Hauteville H, Sansonetti PJ, Carlier MF & Cossart P (1995) The unrelated surface proteins ActA of Listeria monocytogenes and IcsA of Shigella flexneri are sufficient to confer actin-based motility on Listeria innocua and Escherichia coli respectively. Mol. Microbiol. 18: 413–423Google Scholar
  22. LaBrec EH, Schneider H, Magnani TJ & Formal SB (1964) Epithelial cell penetration as an essential step in the pathogenesis of bacillary dysentery. J. Bacteriol. 88: 1503–1518Google Scholar
  23. Makino S, Sasakawa C, Kamata K, Kurata T & Yoshikawa M (1986) A virulence determinant required for continuous reinfection of adjacent cells on large plasmic in Shigella flexneri 2a. Cell 46: 551–555Google Scholar
  24. Maurelli AT, Baudry B, d & #x2019;Hauteville H, Hale TL & Sansonetti PJ (1985) Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexneri. Infect. Immun. 49: 164–171Google Scholar
  25. M & #x00E9;nard R, Sansonetti PJ & Parsot C (1993) Non polar mutagenesis of the ipa genes defines IpaB, IpaC and IpaD as effectors of Shigella flexneri entry into epithelial cells. J. Bacteriol. 175: 5899–5906Google Scholar
  26. M & #x00E9;nard R, Sansonetti PJ, Parsot C & Vasselon T (1994) Extracellular association and cytoplasmic partitioning of the IpaB and IpaC invasins of Shigella flexneri. Cell 79: 515–525Google Scholar
  27. M & #x00E9;nard R, Sansonetti PJ & Parsot C (1994) The secretion of the Shigella flexneri Ipa invasins is induced by the epithelial cell and controlled by IpaB and IpaD. EMBO J. 13: 5293–5302Google Scholar
  28. M & #x00E9;nard R, Pr & #x00E9;vost MC, Gounon P, Sansonetti PJ & Dehio C (1996) The secreted Ipa complex of Shigella flexneri promotes entry into mammalian cells. Proc. Natl. Acad. Sci. USA 93: 1254–1258Google Scholar
  29. M & #x00E9;nard R, Dehio C & Sansonetti PJ (1996) Bacterial entry into epithelial cells: the paradigm of Shigella. Trends Microbiol. 4: 220–226Google Scholar
  30. Mengaud J, Ohayon H, Gounon P, Mège R-M & Cossart P (1996) E-cadherin is the receptor for internalin, a surface protein required for entry of Listeria monocytogenes into epithelial cells. Cell 84: 923–932Google Scholar
  31. Niebuhr K, Chakraborty T, Rohde M, Gazlig T, Jansen B, Kollner P & Wehland J (1993) Localization of the ActA polypeptide of Listeria monocytogenes in infected tissue culture cell lines: ActA is not associated with actin & #x2019;comets & #x2019;. Infect. Immun. 61: 2793–2802Google Scholar
  32. Pr & #x00E9;vost MC, Lesourd M, Arpin M, Vernel F. Mounier J, Hellio R & Sansonetti PJ (1992) Unipolar reorganization of F-actin layer at bacterial division and bundling of actin filaments by plastin correlate with movement of Shigella flexneri within HeLa cells. Infect. Immun. 60: 4088–4099Google Scholar
  33. Reinhard M, Rudiger M, Jockuseh BM & Walter U (1996) VASP interaction with vinculin: a recurring theme of interactions with proline-rich motifs. FEBS Lett. 399: 103–107Google Scholar
  34. Sansonetti PJ, Kopecko DJ & Formal SB (1982) Involvement of a large plasmid in the invasive ability of Shigella flexneri. Infect. Immun. 35: 852–860Google Scholar
  35. Sansonetti PJ, Hale TL, Dammin GJ, Kapfer C, Colins H & Formal S (1983) Alterations in the pathogenicity of Escherichia coli K-12 after transfer of plasmid and chromosomal genes from Shigella flexneri. Infect. Immun. 39: 1392–1402Google Scholar
  36. Sansonetti PJ, Ryter A, Clerc P, Maurelli AT & Mounier J (1986) Multiplication of Shigella flexneri within HeLa cells: lysis of the phagocytic vacuole and plasmid-mediated contact hemolysis. Infect. Immun. 51: 461–469Google Scholar
  37. Sansonetti PJ, Arondel J, Fontaine C, d & #x2019;Hauteville H & Bernardini ML (1991) OmpB (osmo-regulation) and icsA (cell to cell spread) mutants of Shigella flexneri. Evaluation as vaccine candidates. Probes to study the pathogenesis of shigellosis. Vaccine 9: 416–422Google Scholar
  38. Sansonetti PJ, Mounier J, Pr & #x00E9;vost MC & Mège RM (1994) Cadherin expression is required for the spread of Shigella flexneri between epithelial cells. Cell 76: 829–839Google Scholar
  39. Suzuki T, Shinsuke S & Sasakawa C (1996) Functional analysis of Shigella VirG domains essential for interaction with vinculin and actin-based motility. J. Biol. Chem. 271: 21878–21885Google Scholar
  40. Tran Van Nhieu G & Isberg RR (1993) Affinity and receptor density are primary determinants of & #x03B2;1 chain integrin-mediated bacterial internalization. EMBO J. 12: 1887–1895Google Scholar
  41. Tran Van Nhieu G, Ben Ze & #x2019;ev A & Sansonetti PJ (1997) Modulation of bacterial entry in epithelial cells by association between vinculin and the Shigella IpaA invasin. EMBO J. 16: 2717–2729Google Scholar
  42. Vasselon T, Mounier J, Hellio R & Sansonetti PJ (1992) Movement along actin filaments of the perijunctional area and de novo polymerization of cellular actin are required for Shigella flexneri colonization of epithelial Caco-2 cell monolayers. Infect. Immun. 60: 1031–1040Google Scholar
  43. Watarai M, Funato S & Sasakawa C (1996) Interaction of Ipa proteins of Shigella flexneri with alpha5 beta1 integrin promotes entry of the bacteria into mammalian cells. J. Exp. Med. 183: 991–999Google Scholar
  44. Watarai M, Kamata Y, Kozaki S & Sasakawa C (1997) rho, a small GTP-binding protein, is essential for Shigella invasion of epithelial cells. J. Exp. Med. 185: 281–292Google Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • Philippe J. Sansonetti
    • 1
  • Coumaran Egile
    • 1
  1. 1.Unité de Pathogénie Microbienne Moléculaire, INSERM U 389, Institut PasteurParis Cédex 15France

Personalised recommendations