Search tips
Search criteria 


Logo of iaiPermissionsJournals.ASM.orgJournalIAI ArticleJournal InfoAuthorsReviewers
Infect Immun. 1997 December; 65(12): 5309–5319.
PMCID: PMC175764

Internalin of Listeria monocytogenes with an intact leucine-rich repeat region is sufficient to promote internalization.


Listeria monocytogenes can use two different surface proteins, internalin (InlA) and InlB, to invade mammalian cells. The exact role of these invasiveness factors in vivo remains to be determined. In cultured cells, InlA is necessary to promote Listeria entry into human epithelial cells, such as Caco-2 cells, whereas InlB is necessary to promote Listeria internalization in several other cell types, including hepatocytes, fibroblasts, and epithelioid cells, such as Vero, HeLa, CHO, or Hep-2 cells. We have recently reported that the InlA receptor on Caco-2 cells is the cell adhesion molecule E-cadherin and demonstrated that nonpermissive fibroblasts become permissive for internalin-mediated entry when transfected with the gene coding for LCAM, the chicken homolog of the human E-cadherin gene. In this study, we demonstrate for the first time that the internalin protein alone is sufficient to promote internalization into cells expressing its receptor. Indeed, internalin confers invasiveness to both Enterococcus faecalis and internalin-coated latex beads. As shown by transmission electron microscopy, these beads were phagocytosed via a "zipper" mechanism similar to that observed during the internalin-E-cadherin-mediated entry of Listeria. Moreover, a functional analysis of internalin demonstrates that its amino-terminal region, encompassing the leucine-rich repeat (LRR) region and the inter-repeat (IR) region, is necessary and sufficient to promote bacterial entry into cells expressing its receptor. Several lines of evidence suggest that the LRR region would interact directly with E-cadherin, whereas the IR region would be required for a proper folding of the LRR region.

Full Text

The Full Text of this article is available as a PDF (1.2M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Braun L, Dramsi S, Dehoux P, Bierne H, Lindahl G, Cossart P. InlB: an invasion protein of Listeria monocytogenes with a novel type of surface association. Mol Microbiol. 1997 Jul;25(2):285–294. [PubMed]
  • Brückner R. A series of shuttle vectors for Bacillus subtilis and Escherichia coli. Gene. 1992 Dec 1;122(1):187–192. [PubMed]
  • Casadaban MJ, Cohen SN. Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol. 1980 Apr;138(2):179–207. [PubMed]
  • Chen H, Paradies NE, Fedor-Chaiken M, Brackenbury R. E-cadherin mediates adhesion and suppresses cell motility via distinct mechanisms. J Cell Sci. 1997 Feb;110(Pt 3):345–356. [PubMed]
  • Cossart P. Host/pathogen interactions. Subversion of the mammalian cell cytoskeleton by invasive bacteria. J Clin Invest. 1997 May 15;99(10):2307–2311. [PMC free article] [PubMed]
  • Cruz-Rodz AL, Gilmore MS. High efficiency introduction of plasmid DNA into glycine treated Enterococcus faecalis by electroporation. Mol Gen Genet. 1990 Oct;224(1):152–154. [PubMed]
  • Dramsi S, Biswas I, Maguin E, Braun L, Mastroeni P, Cossart P. Entry of Listeria monocytogenes into hepatocytes requires expression of inIB, a surface protein of the internalin multigene family. Mol Microbiol. 1995 Apr;16(2):251–261. [PubMed]
  • Dramsi S, Dehoux P, Lebrun M, Goossens PL, Cossart P. Identification of four new members of the internalin multigene family of Listeria monocytogenes EGD. Infect Immun. 1997 May;65(5):1615–1625. [PMC free article] [PubMed]
  • el Tayar N. Advances in the molecular understanding of gonadotropins-receptors interactions. Mol Cell Endocrinol. 1996 Dec 20;125(1-2):65–70. [PubMed]
  • Finlay BB, Cossart P. Exploitation of mammalian host cell functions by bacterial pathogens. Science. 1997 May 2;276(5313):718–725. [PubMed]
  • Gaillard JL, Berche P, Frehel C, Gouin E, Cossart P. Entry of L. monocytogenes into cells is mediated by internalin, a repeat protein reminiscent of surface antigens from gram-positive cocci. Cell. 1991 Jun 28;65(7):1127–1141. [PubMed]
  • Gaillard JL, Berche P, Mounier J, Richard S, Sansonetti P. In vitro model of penetration and intracellular growth of Listeria monocytogenes in the human enterocyte-like cell line Caco-2. Infect Immun. 1987 Nov;55(11):2822–2829. [PMC free article] [PubMed]
  • Gaillard JL, Finlay BB. Effect of cell polarization and differentiation on entry of Listeria monocytogenes into the enterocyte-like Caco-2 cell line. Infect Immun. 1996 Apr;64(4):1299–1308. [PMC free article] [PubMed]
  • Ireton K, Cossart P. Host-pathogen interactions during entry and actin-based movement of Listeria monocytogenes. Annu Rev Genet. 1997;31:113–138. [PubMed]
  • Ireton K, Payrastre B, Chap H, Ogawa W, Sakaue H, Kasuga M, Cossart P. A role for phosphoinositide 3-kinase in bacterial invasion. Science. 1996 Nov 1;274(5288):780–782. [PubMed]
  • Isberg RR, Leong JM. Multiple beta 1 chain integrins are receptors for invasin, a protein that promotes bacterial penetration into mammalian cells. Cell. 1990 Mar 9;60(5):861–871. [PubMed]
  • Isberg RR, Van Nhieu GT. Two mammalian cell internalization strategies used by pathogenic bacteria. Annu Rev Genet. 1994;28:395–422. [PubMed]
  • Isberg RR, Voorhis DL, Falkow S. Identification of invasin: a protein that allows enteric bacteria to penetrate cultured mammalian cells. Cell. 1987 Aug 28;50(5):769–778. [PubMed]
  • Kobe B, Deisenhofer J. The leucine-rich repeat: a versatile binding motif. Trends Biochem Sci. 1994 Oct;19(10):415–421. [PubMed]
  • Lasa I, David V, Gouin E, Marchand JB, Cossart P. The amino-terminal part of ActA is critical for the actin-based motility of Listeria monocytogenes; the central proline-rich region acts as a stimulator. Mol Microbiol. 1995 Nov;18(3):425–436. [PubMed]
  • Lebrun M, Mengaud J, Ohayon H, Nato F, Cossart P. Internalin must be on the bacterial surface to mediate entry of Listeria monocytogenes into epithelial cells. Mol Microbiol. 1996 Aug;21(3):579–592. [PubMed]
  • Lee FS, Vallee BL. Modular mutagenesis of human placental ribonuclease inhibitor, a protein with leucine-rich repeats. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1879–1883. [PubMed]
  • Leong JM, Morrissey PE, Isberg RR. A 76-amino acid disulfide loop in the Yersinia pseudotuberculosis invasin protein is required for integrin receptor recognition. J Biol Chem. 1993 Sep 25;268(27):20524–20532. [PubMed]
  • Lorber B. Listeriosis. Clin Infect Dis. 1997 Jan;24(1):1–11. [PubMed]
  • Marra A, Isberg RR. Common entry mechanisms. Bacterial pathogenesis. Curr Biol. 1996 Sep 1;6(9):1084–1086. [PubMed]
  • Mengaud J, Lecuit M, Lebrun M, Nato F, Mazie JC, Cossart P. Antibodies to the leucine-rich repeat region of internalin block entry of Listeria monocytogenes into cells expressing E-cadherin. Infect Immun. 1996 Dec;64(12):5430–5433. [PMC free article] [PubMed]
  • 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. [PubMed]
  • Saltman LH, Lu Y, Zaharias EM, Isberg RR. A region of the Yersinia pseudotuberculosis invasin protein that contributes to high affinity binding to integrin receptors. J Biol Chem. 1996 Sep 20;271(38):23438–23444. [PubMed]
  • Schneewind O, Mihaylova-Petkov D, Model P. Cell wall sorting signals in surface proteins of gram-positive bacteria. EMBO J. 1993 Dec;12(12):4803–4811. [PubMed]
  • Schneewind O, Model P, Fischetti VA. Sorting of protein A to the staphylococcal cell wall. Cell. 1992 Jul 24;70(2):267–281. [PubMed]
  • Swanson JA, Baer SC. Phagocytosis by zippers and triggers. Trends Cell Biol. 1995 Mar;5(3):89–93. [PubMed]
  • Trieu-Cuot P, Carlier C, Poyart-Salmeron C, Courvalin P. Shuttle vectors containing a multiple cloning site and a lacZ alpha gene for conjugal transfer of DNA from Escherichia coli to gram-positive bacteria. Gene. 1991 Jun 15;102(1):99–104. [PubMed]
  • Van Putten JP, Weel JF, Grassmé HU. Measurements of invasion by antibody labeling and electron microscopy. Methods Enzymol. 1994;236:420–437. [PubMed]
  • Ware J, Russell SR, Marchese P, Murata M, Mazzucato M, De Marco L, Ruggeri ZM. Point mutation in a leucine-rich repeat of platelet glycoprotein Ib alpha resulting in the Bernard-Soulier syndrome. J Clin Invest. 1993 Sep;92(3):1213–1220. [PMC free article] [PubMed]
  • Yanisch-Perron C, Vieira J, Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. [PubMed]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)