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. 1997 May 15;16(10):2717–2729. doi: 10.1093/emboj/16.10.2717

Modulation of bacterial entry into epithelial cells by association between vinculin and the Shigella IpaA invasin.

G Tran Van Nhieu 1, A Ben-Ze'ev 1, P J Sansonetti 1
PMCID: PMC1169882  PMID: 9184218

Abstract

Shigella flexneri is the causative agent of bacillary dysentery in humans. Shigella invasion of epithelial cells is characterized by cytoskeletal rearrangements and formation of cellular projections engulfing the bacterium in a macropinocytic process. We show here that vinculin, a protein involved in linking actin filaments to the plasma membrane, is a direct target of Shigella during cell invasion. IpaA, a Shigella protein secreted upon cell contact, rapidly associates with vinculin during bacterial invasion. Although defective for cell entry, an ipaA mutant is still able to induce foci of actin polymerization, but differs from wild-type Shigella in its ability to recruit vinculin and alpha-actinin. Presumably, IpaA-vinculin interaction initiates the formation of focal adhesion-like structures required for efficient invasion.

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Selected References

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  1. Adam T., Arpin M., Prévost M. C., Gounon P., Sansonetti P. J. Cytoskeletal rearrangements and the functional role of T-plastin during entry of Shigella flexneri into HeLa cells. J Cell Biol. 1995 Apr;129(2):367–381. doi: 10.1083/jcb.129.2.367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ben-Ze'ev A., Reiss R., Bendori R., Gorodecki B. Transient induction of vinculin gene expression in 3T3 fibroblasts stimulated by serum-growth factors. Cell Regul. 1990 Aug;1(9):621–636. doi: 10.1091/mbc.1.9.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernardini M. L., Mounier J., d'Hauteville H., Coquis-Rondon M., Sansonetti P. J. Identification of icsA, a plasmid locus of Shigella flexneri that governs bacterial intra- and intercellular spread through interaction with F-actin. Proc Natl Acad Sci U S A. 1989 May;86(10):3867–3871. doi: 10.1073/pnas.86.10.3867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brasaemle D. L., Attie A. D. Microelisa reader quantitation of fixed, stained, solubilized cells in microtitre dishes. Biotechniques. 1988 May;6(5):418–419. [PubMed] [Google Scholar]
  5. Craig S. W., Johnson R. P. Assembly of focal adhesions: progress, paradigms, and portents. Curr Opin Cell Biol. 1996 Feb;8(1):74–85. doi: 10.1016/s0955-0674(96)80051-2. [DOI] [PubMed] [Google Scholar]
  6. Finlay B. B., Ruschkowski S., Dedhar S. Cytoskeletal rearrangements accompanying salmonella entry into epithelial cells. J Cell Sci. 1991 Jun;99(Pt 2):283–296. doi: 10.1242/jcs.99.2.283. [DOI] [PubMed] [Google Scholar]
  7. Francis C. L., Ryan T. A., Jones B. D., Smith S. J., Falkow S. Ruffles induced by Salmonella and other stimuli direct macropinocytosis of bacteria. Nature. 1993 Aug 12;364(6438):639–642. doi: 10.1038/364639a0. [DOI] [PubMed] [Google Scholar]
  8. Groisman E. A., Ochman H. Cognate gene clusters govern invasion of host epithelial cells by Salmonella typhimurium and Shigella flexneri. EMBO J. 1993 Oct;12(10):3779–3787. doi: 10.1002/j.1460-2075.1993.tb06056.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hermant D., Ménard R., Arricau N., Parsot C., Popoff M. Y. Functional conservation of the Salmonella and Shigella effectors of entry into epithelial cells. Mol Microbiol. 1995 Aug;17(4):781–789. doi: 10.1111/j.1365-2958.1995.mmi_17040781.x. [DOI] [PubMed] [Google Scholar]
  10. Isberg R. R. Discrimination between intracellular uptake and surface adhesion of bacterial pathogens. Science. 1991 May 17;252(5008):934–938. doi: 10.1126/science.1674624. [DOI] [PubMed] [Google Scholar]
  11. Isberg R. R., Falkow S. A single genetic locus encoded by Yersinia pseudotuberculosis permits invasion of cultured animal cells by Escherichia coli K-12. Nature. 1985 Sep 19;317(6034):262–264. doi: 10.1038/317262a0. [DOI] [PubMed] [Google Scholar]
  12. Isberg R. R., Van Nhieu G. T. Two mammalian cell internalization strategies used by pathogenic bacteria. Annu Rev Genet. 1994;28:395–422. doi: 10.1146/annurev.ge.28.120194.002143. [DOI] [PubMed] [Google Scholar]
  13. Johnson R. P., Craig S. W. F-actin binding site masked by the intramolecular association of vinculin head and tail domains. Nature. 1995 Jan 19;373(6511):261–264. doi: 10.1038/373261a0. [DOI] [PubMed] [Google Scholar]
  14. Kadurugamuwa J. L., Rohde M., Wehland J., Timmis K. N. Intercellular spread of Shigella flexneri through a monolayer mediated by membranous protrusions and associated with reorganization of the cytoskeletal protein vinculin. Infect Immun. 1991 Oct;59(10):3463–3471. doi: 10.1128/iai.59.10.3463-3471.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kaniga K., Trollinger D., Galán J. E. Identification of two targets of the type III protein secretion system encoded by the inv and spa loci of Salmonella typhimurium that have homology to the Shigella IpaD and IpaA proteins. J Bacteriol. 1995 Dec;177(24):7078–7085. doi: 10.1128/jb.177.24.7078-7085.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lee C., Levin A., Branton D. Copper staining: a five-minute protein stain for sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1987 Nov 1;166(2):308–312. doi: 10.1016/0003-2697(87)90579-3. [DOI] [PubMed] [Google Scholar]
  18. Maurelli A. T., Baudry B., d'Hauteville H., Hale T. L., Sansonetti P. J. Cloning of plasmid DNA sequences involved in invasion of HeLa cells by Shigella flexneri. Infect Immun. 1985 Jul;49(1):164–171. doi: 10.1128/iai.49.1.164-171.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mengaud J., Ohayon H., Gounon P., Mege R-M, Cossart P. E-cadherin is the receptor for internalin, a surface protein required for entry of L. monocytogenes into epithelial cells. Cell. 1996 Mar 22;84(6):923–932. doi: 10.1016/s0092-8674(00)81070-3. [DOI] [PubMed] [Google Scholar]
  20. Menkel A. R., Kroemker M., Bubeck P., Ronsiek M., Nikolai G., Jockusch B. M. Characterization of an F-actin-binding domain in the cytoskeletal protein vinculin. J Cell Biol. 1994 Sep;126(5):1231–1240. doi: 10.1083/jcb.126.5.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ménard R., Sansonetti P. J., Parsot C. Nonpolar mutagenesis of the ipa genes defines IpaB, IpaC, and IpaD as effectors of Shigella flexneri entry into epithelial cells. J Bacteriol. 1993 Sep;175(18):5899–5906. doi: 10.1128/jb.175.18.5899-5906.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nobes C. D., Hall A. Rho, rac, and cdc42 GTPases regulate the assembly of multimolecular focal complexes associated with actin stress fibers, lamellipodia, and filopodia. Cell. 1995 Apr 7;81(1):53–62. doi: 10.1016/0092-8674(95)90370-4. [DOI] [PubMed] [Google Scholar]
  23. Parsons J. T. Integrin-mediated signalling: regulation by protein tyrosine kinases and small GTP-binding proteins. Curr Opin Cell Biol. 1996 Apr;8(2):146–152. doi: 10.1016/s0955-0674(96)80059-7. [DOI] [PubMed] [Google Scholar]
  24. Parsot C., Ménard R., Gounon P., Sansonetti P. J. Enhanced secretion through the Shigella flexneri Mxi-Spa translocon leads to assembly of extracellular proteins into macromolecular structures. Mol Microbiol. 1995 Apr;16(2):291–300. doi: 10.1111/j.1365-2958.1995.tb02301.x. [DOI] [PubMed] [Google Scholar]
  25. Rodríguez Fernández J. L., Geiger B., Salomon D., Sabanay I., Zöller M., Ben-Ze'ev A. Suppression of tumorigenicity in transformed cells after transfection with vinculin cDNA. J Cell Biol. 1992 Oct;119(2):427–438. doi: 10.1083/jcb.119.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rosqvist R., Håkansson S., Forsberg A., Wolf-Watz H. Functional conservation of the secretion and translocation machinery for virulence proteins of yersiniae, salmonellae and shigellae. EMBO J. 1995 Sep 1;14(17):4187–4195. doi: 10.1002/j.1460-2075.1995.tb00092.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rosqvist R., Magnusson K. E., Wolf-Watz H. Target cell contact triggers expression and polarized transfer of Yersinia YopE cytotoxin into mammalian cells. EMBO J. 1994 Feb 15;13(4):964–972. doi: 10.1002/j.1460-2075.1994.tb06341.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sasakawa C., Kamata K., Sakai T., Makino S., Yamada M., Okada N., Yoshikawa M. Virulence-associated genetic regions comprising 31 kilobases of the 230-kilobase plasmid in Shigella flexneri 2a. J Bacteriol. 1988 Jun;170(6):2480–2484. doi: 10.1128/jb.170.6.2480-2484.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Suzuki T., Saga S., Sasakawa C. Functional analysis of Shigella VirG domains essential for interaction with vinculin and actin-based motility. J Biol Chem. 1996 Sep 6;271(36):21878–21885. doi: 10.1074/jbc.271.36.21878. [DOI] [PubMed] [Google Scholar]
  30. Tran Van Nhieu G., Isberg R. R. Bacterial internalization mediated by beta 1 chain integrins is determined by ligand affinity and receptor density. EMBO J. 1993 May;12(5):1887–1895. doi: 10.1002/j.1460-2075.1993.tb05837.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Varnum-Finney B., Reichardt L. F. Vinculin-deficient PC12 cell lines extend unstable lamellipodia and filopodia and have a reduced rate of neurite outgrowth. J Cell Biol. 1994 Nov;127(4):1071–1084. doi: 10.1083/jcb.127.4.1071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Watarai M., Funato S., Sasakawa C. Interaction of Ipa proteins of Shigella flexneri with alpha5beta1 integrin promotes entry of the bacteria into mammalian cells. J Exp Med. 1996 Mar 1;183(3):991–999. doi: 10.1084/jem.183.3.991. [DOI] [PMC free article] [PubMed] [Google Scholar]

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