Skip to main content
Download PDF

Integrin receptors play a role in the internalin B-dependent entry of Listeria monocytogenes into host cells

Cellular & Molecular Biology Letters volume 15, pages 496–506 (2010)Cite this article

Abstract

Listeria monocytogenes enters non-phagocytic cells by binding its surface proteins inlA (internalin) and inlB to the host’s E-cadherin and Met, respectively. The two internalins play either separate or cooperative roles in the colonization of infected tissues. Here, we studied bacterial uptake into HeLa cells using an L. monocytogenes mutant strain (ΔinlA) carrying a deletion in the gene coding for inlA. The ΔinlA mutant strain showed the capability to invade HeLa cells. The monoclonal anti-β3- and anti-β1-integrin subunit antibodies prevented bacterial uptake into the cells, while the anti-β2- and anti-β4-integrin subunit antibodies failed to affect L. monocytogenes entry into HeLa cells. Three structurally distinct disintegrins (kistrin, echistatin and flavoridin) also inhibited bacterial uptake, showing different potencies correlated to their selective affinity for the β3- and β1-integrin subunits. In addition to inducing Met phosphorylation, infection of cells by the L. monocytogenes ΔinlA mutant strain promoted the tyrosine phosphorylation of the focal adhesion-associated proteins FAK and paxillin. Our findings provide the first evidence that β3- and β1-integrin receptors play a role in the inlB-dependent internalization of L. monocytogenes into host cells.

Abbreviations

BHI:

brain-heart infusion

BSA:

bovine serum albumin

CFU:

colony-forming unit

DMEM:

Dulbecco’s modified Eagle’s medium

FAK:

focal adhesion kinase

FBS:

bovine fetal serum

InlA:

internalin A

InlB:

internalin B

MOI:

multiplicity of infection

PBS:

phosphate-buffered saline

SDS:

sodium dodecyl sulphate

References

  1. Hamon, M., Bierne, H. and Cossart, P. Listeria monocytogenes: a multifaceted model. Nat. Rev. Microbiol. 4 (2006) 423–434.

    Article  CAS  PubMed  Google Scholar 

  2. Ireton, K. Entry of the bacterial pathogen Listeria monocytogenes into mammalian cells. Cell. Microbiol. 9 (2997) 1365–1375.

    Article  Google Scholar 

  3. Bierne, H., Sabet, C., Personnic, N. and Cossart, P. Internalins: a complex family of leucine-rich repeat-containing proteins in Listeria monocytogenes. Microbes Infect. 9 (2007) 1156–1166.

    Article  CAS  PubMed  Google Scholar 

  4. Mengaud, J., Ohayon, H., Gounon, P., Mege, R-M. and Cossart P. E-cadherin is the receptor for internalin, a surface protein required for entry of L. monocytogenes into epithelial cells. Cell 84 (1996) 923–932.

    Article  CAS  PubMed  Google Scholar 

  5. Bonazzi, M., Lecuit, M. and Cossart, P. Listeria monocytogenes internalin and E-cadherin: from structure to pathogenesis. Cell. Microbiol. 11 (2009) 693–702.

    Article  CAS  Google Scholar 

  6. Shen, Y., Naujokas, M., Park, M. and Ireton K. InIB-dependent internalization of Listeria is mediated by the Met receptor tyrosine kinase. Cell 103 (2000) 501–510.

    Article  CAS  PubMed  Google Scholar 

  7. Pizarro-Cerdá, J., Sousa, S. and Cossart, P. Exploitation of host cell cytoskeleton and signalling during Listeria monocytogenes entry into mammalian cells. C. R. Biol. 327 (2004) 115–123.

    Article  PubMed  Google Scholar 

  8. Cossart, P. Met, the HGF-SF receptor: another receptor for Listeria monocytogenes. Trends Microbiol. 9 (2001) 105–107.

    Article  CAS  PubMed  Google Scholar 

  9. Eliceiri, B.P. Integrin and growth factor receptor crosstalk. Circ. Res. 89 (2001) 1104–1110.

    Article  CAS  PubMed  Google Scholar 

  10. Trusolino, L., Bertotti, A. and Comoglio P.M. A signaling adapter function for alpha6beta4 integrin in the control of HGF-dependent invasive growth. Cell 107 (2001) 643–654.

    Article  CAS  PubMed  Google Scholar 

  11. Streuli, C.H. and Akhtar, N. Signal co-operation between integrins and other receptor systems. Biochem. J. 418 (2009) 491–506.

    Article  CAS  PubMed  Google Scholar 

  12. Hynes, R.O. Integrins: bidirectional, allosteric signaling machines. Cell 110 (2002) 673–687.

    Article  CAS  PubMed  Google Scholar 

  13. Sastry, S.K. and Burridge, K. Focal adhesions: a nexus for intracellular signaling and cytoskeletal dynamics. Exp. Cell Res. 261 (2000) 25–36.

    Article  CAS  PubMed  Google Scholar 

  14. Dupuy, A.G. and Caron, E. Integrin-dependent phagocytosis: spreading from microadhesion to new concepts. J. Cell Sci. 121 (2008) 1773–1783.

    Article  CAS  PubMed  Google Scholar 

  15. Vicente-Manzanares, M., Choi, C.K. and Horwitz, A.R. Integrins in cell migration-the actin connection. J. Cell Sci. 122 (2009) 199–206.

    Article  CAS  PubMed  Google Scholar 

  16. Dersch, P. Molecular and cellular mechanisms of bacterial entry into host cells. Contrib. Microbiol. 10 (2003) 183–209.

    Article  CAS  PubMed  Google Scholar 

  17. Scibelli, A., Roperto, S., Manna, L., Pavone, L.M., Tafuri, S., Della Morte, R. and Staiano, N. Engagement of integrins as a cellular route of invasion by bacterial pathogens. Vet. J. 173 (2007) 482–491.

    Article  CAS  PubMed  Google Scholar 

  18. Kwok, T., Zabler, D., Urman, S., Rohde, M., Hartig, R., Wessler, S., Misselwitz, R., Berger, J., Sewald, N., König, W. and Backert, S. Helicobacter exploits integrin for type IV secretion and kinase activation. Nature 449 (2007) 862–866.

    Article  CAS  PubMed  Google Scholar 

  19. Bergmann, B., Raffelsbauer, D., Kuhn, M., Goetz, M., Hom, S. and Goebel, W. InlA- but not InlB-mediated internalization of Listeria monocytogenes by non-phagocytic mammalian cells needs the support of other internalins. Mol. Microbiol. 43 (2002) 557–570.

    Article  CAS  PubMed  Google Scholar 

  20. McLane, M.A., Marcinkiewicz, C., Vijay-Kumar, S., Wierzbicka-Patynowski, I. and Niewiarowski, S. Viper venom disintegrins and related molecules. Proc. Soc. Exp. Biol. Med. 219 (1998) 109–119.

    CAS  PubMed  Google Scholar 

  21. Scibelli, A., Matteoli, G., Roperto, S., Alimenti, E., Dipineto, L., Pavone, L.M., Della Morte, R., Menna, L.F., Fioretti, A. and Staiano, N. Flavoridin inhibits Yersinia enterocolitica uptake into fibronectin-adherent HeLa cells. FEMS Microbiol. Lett. 247 (2005) 51–57.

    Article  CAS  PubMed  Google Scholar 

  22. Lu, X., Lu, D., Scully, M.F. and Kakkar, V.V. Modulation of integrin-binding selectivity by mutation within the RGD-loop of snake venom proteins: a novel drug development approach. Curr. Med. Chem. Cardiovasc. Hematol. Agents 1 (2003) 89–196.

    Article  Google Scholar 

  23. Senn, H. and Klaus, W. The nuclear magnetic resonance solution structure of flavoridin, an antagonist of the platelet GP IIb-IIIa receptor. J. Mol. Biol. 232 (1993) 907–925.

    Article  CAS  PubMed  Google Scholar 

  24. Schaller, M.D. Biochemical signals and biological responses elicited by the focal adhesion kinase. Biochim. Biophys. Acta 1540 (2001) 1–21.

    Article  CAS  PubMed  Google Scholar 

  25. Hamadi, A., Bouali, M., Dontenwill, M., Stoeckel, H., Takeda, K. and Rondé, P. Regulation of focal adhesion dynamics and disassembly by phosphorylation of FAK at tyrosine 397. J. Cell Sci. 118 (2005) 4415–4425.

    Article  CAS  PubMed  Google Scholar 

  26. Staiano, N., Garbi, C., Squillacioti, C., Esposito, S., Di Martino, E., Belisario, M.A., Nitsch, L. and Di Natale, P. Echistatin induces decrease of pp125FAK phosphorylation, disassembly of actin cytoskeleton and focal adhesions, and detachment of fibronectin-adherent melanoma cells. Eur. J. Cell Biol. 73 (1997) 298–305.

    CAS  PubMed  Google Scholar 

  27. Della Morte, R., Squillacioti, C., Garbi, C., Derkinderen, P., Belisario, M.A., Girault, J.A., Di Natale, P., Nitsch, L. and Staiano, N. Echistatin inhibits pp125FAK autophosphorylation, paxillin phosphorylation and pp125FAK-paxillin interaction in fibronectin-adherent melanoma cells. Eur. J. Biochem. 267 (2000) 5047–5054.

    Article  CAS  PubMed  Google Scholar 

  28. van Nimwegen, M.J. and van de Water, B. Focal adhesion kinase: a potential target in cancer therapy. Biochem. Pharmacol. 73 (2007) 597–609.

    Article  PubMed  Google Scholar 

  29. McCall-Culbreath, K. D., Li, Z. and Zutter, M.M. Crosstalk between the alpha2beta1 integrin and c-met/HGF-R regulates innate immunity. Blood 111 (2008) 3562–3570.

    Article  CAS  PubMed  Google Scholar 

  30. Cabodi, S., Moro, L., Bergatto, E., Boeri Erba, E., Di Stefano, P., Turco, E., Tarone, G. and Defilippi, P. Integrin regulation of epidermal growth factor (EGF) receptor and of EGF-dependent responses. Biochem. Soc. Trans. 32 (2004) 438–442.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Istituto Zooprofilattico Sperimentale del Mezzogiorno, via Della Salute 2, 80055, Portici (Na), Italy

    Clementina Auriemma, Maurizio Viscardi, Federico Capuano & Giuseppe Iovane

  2. Dipartimento di Strutture, Funzioni e Tecnologie Biologiche, Università degli Studi di Napoli Federico II, via F. Delpino 1, 80137, Napoli, Italy

    Simona Tafuri, Luigi Michele Pavone, Rossella Della Morte & Norma Staiano

  3. Dipartimento di Patologia e Sanità Animale, Università degli Studi di Napoli Federico II, via F. Delpino 1, 80137, Napoli, Italy

    Laura Rinaldi & Giuseppe Iovane

Authors
  1. Clementina Auriemma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maurizio Viscardi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Simona Tafuri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Luigi Michele Pavone
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Federico Capuano
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Laura Rinaldi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Rossella Della Morte
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Giuseppe Iovane
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Norma Staiano
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Norma Staiano.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Auriemma, C., Viscardi, M., Tafuri, S. et al. Integrin receptors play a role in the internalin B-dependent entry of Listeria monocytogenes into host cells. Cell Mol Biol Lett 15, 496–506 (2010). https://doi.org/10.2478/s11658-010-0019-z

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.2478/s11658-010-0019-z

Key words

  • Listeria monocytogenes
  • Internalin B
  • Integrins
  • FAK
  • Paxillin

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us