[en] Enteropathogenic Escherichia coli (EPEC) and enterohaemorrhagic E. coli (EHEC) are closely related pathogens. Both use a type III secretion system (TTSS) encoded by the 'locus of enterocyte effacement' (LEE) to subvert and attach to epithelial cells through the injection of a repertoire of effector molecules. Here, we report the identification of a new TTSS translocated effector molecule called Cif, which blocks cell cycle G(2)/M transition and induces the formation of stress fibres through the recruitment of focal adhesions. Cif is not encoded by the LEE but by a lambdoid prophage present in EPEC and EHEC. A cif mutant causes localized effacement of microvilli and intimately attaches to the host cell surface, but is defective in the ability to block mitosis. When expressed in TTSS competent LEE-positive pathogens, Cif is injected into the infected epithelial cells. These cells arrested at the G(2)/M phase displayed accumulation of inactive phosphorylated Cdk1. In conclusion, Cif is a new member of a growing family of bacterial cyclomodulins that subvert the host eukaryotic cell cycle.
Mainil, Jacques ; Université de Liège - ULiège > Département des maladies infectieuses et parasitaires > Bactériologie et pathologie des maladies bactériennes
Rosenshine, I.
Sugai, M.
De Rycke, J.
Oswald, E.
Language :
English
Title :
Enteropathogenic and enterohaemorrhagic Escherichia coli deliver a novel effector called Cif, which blocks cell cycle G(2)/M transition
Aragon, V., Chao, K., and Dreyfus, L.A. (1997) Effect of cytolethal distending toxin on F-actin assembly and cell division in Chinese hamster ovary cells. Infect Immun 65: 3774-3780.
Blattner, F.R., Plunkett, G., III, Bloch, C.A., Perna, N.T., Burland, V., Riley, M., et al. (1997) The complete genome sequence of Escherichia coli K-12. Science 277: 1453-1474.
Comayras, C., Tasca, C., Peres, S.Y., Ducommun, B., Oswald, E., and De Rycke, J. (1997) Escherichia coli cytolethal distending toxin blocks the HeLa cell cycle at the G2/M transition by preventing cdc2 protein kinase dephosphorylation and activation. Infect Immun 65: 5088-5095.
Cortes-Bratti, X., Chaves-Olarte, E., Lagergard, T., and Thelestam, M. (1999) The cytolethal distending toxin from the chancroid bacterium Haemophilus ducreyi induces cell-cycle arrest in the G2 phase. J Clin Invest 103: 107-115.
Crane, J.K., McNamara, B.P., and Donnenberg, M.S. (2001) Role of EspF in host cell death induced by enteropathogenic Escherichia coli. Cell Microbiol 3: 197-211.
Datsenko, K.A., and Wanner, B.L. (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97: 6640-6645.
De Rycke, J., Comtet, E., Chalareng, C., Boury, M., Tasca, C., and Milon, A. (1997) Enteropathogenic Escherichia coli O103 from rabbit elicits actin stress fibers and focal adhesions in HeLa epithelial cells, cytopathic effects that are linked to an analog of the locus of enterocyte effacement. Infect Immun 65: 2555-2563.
Deibel, C., Kramer, S., Chakraborty, T., and Ebel, F. (1998) EspE, a novel secreted protein of attaching and effacing bacteria, is directly translocated into infected host cells, where it appears as a tyrosine-phosphorylated 90 kDa protein. Mol Microbiol 28: 463-474.
Elliott, S.J., Krejany, E.O., Mellies, J.L., Robins-Browne, R.M., Sasakawa, C., and Kaper, J.B. (2001) EspG, a novel type III system-secreted protein from enteropathogenic Escherichia coli with similarities to VirA of Shigella flexneri. Infect Immun 69: 4027-4033.
Elwell, C.A., and Dreyfus, L.A. (2000) DNase I homologous residues in CdtB are critical for cytolethal distending toxin-mediated cell cycle arrest. Mol Microbiol 37: 952-963.
Frankel, G., Phillips, A.D., Rosenshine, I., Dougan, G., Kaper, J.B., and Knutton, S. (1998) Enteropathogenic and enterohaemorrhagic Escherichia coli: more subversive elements. Mol Microbiol 30: 911-921.
Hayashi, T., Makino, K., Ohnishi, M., Kurokawa, K., Ishii, K., Yokoyama, K., et al. (2001) Complete genome sequence of enterohemorrhagic Escherichia coli O157:H7 and genomic comparison with a laboratory strain K-12. DNA Res 8: 11-22.
Ho, T.D., Figueroa-Bossi, N., Wang, M., Uzzau, S., Bossi, L., and Slauch, J.M. (2002) Identification of GtgE, a novel virulence factor encoded on the Gifsy-2 bacteriophage of Salmonella enterica serovar Typhimurium. J Bacteriol 184: 5234-5239.
Jarvis, K.G., Giron, J.A., Jerse, A.E., McDaniel, T.K., Donnenberg, M.S., and Kaper, J.B. (1995) Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation. Proc Natl Acad Sci USA 92: 7996-8000.
Jerse, A.E., Yu, J., Tall, B.D., and Kaper, J.B. (1990) A genetic locus of enteropathogenic Escherichia coli necessary for the production of attaching and effacing lesions on tissue culture cells. Proc Natl Acad Sci USA 87: 7839-7843.
Kaniga, K., Delor, I., and Cornelis, G.R. (1991) A wide-host-range suicide vector for improving reverse genetics in gram-negative bacteria: inactivation of the blaA gene of Yersinia enterocolitica. Gene 109: 137-141.
Kenny, B., and Jepson, M. (2000) Targeting of an enteropathogenic Escherichia coli (EPEC) effector protein to host mitochondria. Cell Microbiol 2: 579-590.
Kenny, B., DeVinney, R., Stein, M., Reinscheid, D.J., Frey, E.A., and Finlay, B.B. (1997) Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91: 511-520.
Kenny, B., Ellis, S., Leard, A.D., Warawa, J., Mellor, H., and Jepson, M.A. (2002) Co-ordinate regulation of distinct host cell signalling pathways by multifunctional enteropathogenic Escherichia coli effector molecules. Mol Microbiol 44: 1095-1107.
Knutton, S., Baldwin, T., Williams, P.H., and McNeish, A.S. (1989) Actin accumulation at sites of bacterial adhesion to tissue culture cells: basis of a new diagnostic test for enteropathogenic and enterohemorrhagic Escherichia coli. Infect Immun 57: 1290-1298.
Lara-Tejero, M., and Galan, J.E. (2000) A bacterial toxin that controls cell cycle progression as a deoxyribonuclease I-like protein. Science 290: 354-357.
Lara-Tejero, M., and Galan, J.E. (2002) Cytolethal distending toxin: limited damage as a strategy to modulate cellular functions. Trends Microbiol 10: 147-152.
Lax, A.J., and Thomas, W. (2002) How bacteria could cause cancer: one step at a time. Trends Microbiol 10: 293-299.
Li, L., Sharipo, A., Chaves Olarte, E., Masucci, M.G., Levitsky, V., Thelestam, M., and Frisan, T. (2002) The Haemophilus ducreyi cytolethal distending toxin activates sensors of DNA damage and repair complexes in proliferating and non-proliferating cells. Cell Microbiol 4: 87-99.
McDaniel, T.K., Jarvis, K.G., Donnenberg, M.S., and Kaper, J.B. (1995) A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. Proc Natl Acad Sci USA 92: 1664-1668.
McNamara, B.P., Koutsouris, A.O., Connell, C.B., Nougayrede, J.P., Donnenberg, M.S., and Hecht, G. (2001) Translocated EspF protein from enteropathogenic Escherichia coli disrupts host intestinal barrier function. J Clin Invest 107: 621-629.
Marchès, O., Nougayrede, J.P., Boullier, S., Mainil, J., Charlier, G., Raymond, I., et al. (2000) Role of tir and intimin in the virulence of rabbit enteropathogenic Escherichia coli serotype O103:H2. Infect Immun 68: 2171-2182.
Miao, E.A., and Miller, S.I. (2000) A conserved amino acid sequence directing intracellular type III secretion by Salmonella typhimurium. Proc Natl Acad Sci USA 97: 7539-7544.
Mirold, S., Rabsch, W., Rohde, M., Stender, S., Tschape, H., Russmann, H., et al. (1999) Isolation of a temperate bacteriophage encoding the type III effector protein SopE from an epidemic Salmonella typhimurium strain. Proc Natl Acad Sci USA 96: 9845-9850.
Nougayrede, J.P., Marches, O., Boury, M., Mainil, J., Charlier, G., Pohl, P., et al. (1999) The long-term cytoskeletal rearrangement induced by rabbit enteropathogenic Escherichia coli is Esp dependent but intimin independent. Mol Microbiol 31: 19-30.
Nougayrede, J.P., Boury, M., Tasca, C., Marches, O., Milon, A., Oswald, E., and De Rycke, J. (2001) Type III secretion-dependent cell cycle block caused in HeLa cells by enteropathogenic Escherichia coli O103. Infect Immun 69: 6785-6795.
Ohnishi, M., Kurokawa, K., and Hayashi, T. (2001) Diversification of Escherichia coli genomes: are bacteriophages the major contributors? Trends Microbiol 9: 481-485.
Oswald, E., Schmidt, H., Morabito, S., Karch, H., Marches, O., and Caprioli, A. (2000) Typing of intimin genes in human and animal enterohemorrhagic and enteropathogenic Escherichia coli: characterization of a new intimin variant. Infect Immun 68: 64-71.
Pérès, S.Y., Marchès, O., Daigle, F., Nougayrede, J.P., Herault, F., Tasca, C., et al. (1997) A new cytolethal distending toxin (CDT) from Escherichia coli producing CNF2 blocks HeLa cell division in G2/M phase. Mol Microbiol 24: 1095-1107.
Perna, N.T., Plunkett, G., III, Burland, V., Mau, B., Glasner, J.D., Rose, D.J., et al. (2001) Genome sequence of enterohaemorrhagic Escherichia coli O157:H7. Nature 409: 529-533.
Rainbow, L., Hart, C.A., and Winstanley, C. (2002) Distribution of type III secretion gene clusters in Burkholderia pseudomallei, B. thailandensis and B. mallei. J Med Microbiol 51: 374-384.
Salomon, Y. (1991) Cellular responsiveness to hormones and neurotransmitters. conversion of [3H]adenine to [3H]cAMP in cell monolayers, cell suspensions, and tissue slices. Methods Enzymol 195: 22-28.
Stevens, M.P., Wood, M.W., Taylor, L.A., Monaghan, P., Hawes, P., Jones, P.W., et al. (2002) An Inv/Mxi-Spa-like type III protein secretion system in Burkholderia pseudomallei modulates intracellular behaviour of the pathogen. Mol Microbiol 46: 649-659.
Tu, X., Nisan, I., Yona, C., Hanski, E., and Rosenshine, I. (2003) EspH, a new cytoskeleton-modulating effector of enterohaemorrhagic and enteropathogenic Escherichia coli. Mol Microbiol 47: 595-606.
Vallance, B.A., and Finlay, B.B. (2000) Exploitation of host cells by enteropathogenic Escherichia coli. Proc Natl Acad Sci USA 97: 8799-8806.
Wagner, P.L., and Waldor, M.K. (2002) Bacteriophage control of bacterial virulence. Infect Immun 70: 3985-3993.