Summary
Tir of enteropathogenic Escherichia coli (EPEC) or enterohemorrahgic E. coil (EHEC) is translocated by a type III secretion system to the host cell membranes where it serves as a receptor for the binding of a second bacterial membrane protein. In response to the binding, EPEC Tir is phosphorylated at Tyr474, and this phosphorylation is necessary for the signaling of pedestal formation. Tir of EHEC has no equivalent phosphorylation site but it is similarly needed for cytoskeleton rearrangement. How these two Tir molecules achieve their function by apparently different mechanisms is not completely clear. To examine their intrinsic differences, the two Tirs were expressed in HeLa cells and compared. Actin in complexes could be pelleted down from the lysate of cells expressing EHEC Tir but not EPEC Tir. By immunostaining, neither Tir molecule was found in phosphorylated state. In the cytoplasm, EHEC Tir was frequently found in fibrous structures whereas EPEC Tir was observed completely in a diffusive form. The determinant critical for the EHEC Tir fibrous formation was mapped to the C-terminal region of the molecule that deviates from the EPEC counterpart. This region may play a role in taking an alternative route different from Tyr474 phosphorylation to transduce signals.
Article PDF
Similar content being viewed by others
References
Nataro J.P., Kaper J.B. (1998) Diarrheagenic Escherichia coli. Clin Microbiol Rev 11: 142–201
Frankel G., Phillips A.D., Rosenshine I., Dougan G., Kaper J.B., Knutton S. (1998) Enteropathogenic and enterohaemorrhagic Escherichia coli: more subversive elements. Mol Microbiol 30: 911–921
McDaniel T.K., Kaper J.B. (1997) A cloned pathogenicity island from enteropathogenic Escherichia coli confers the attaching and effacing phenotype on E. coli K-12. Mol Microbiol 23:399–407
Perna N.T., Mayhew G.F., Posfai G., Elliott S., Donnenberg M.S., Kaper J.B., Blattner F.R. (1998) Molecular evolution of a pathogenicity island from enterohemorrhagic Escherichia coli O157:H7. Infect Immun 66: 3810–3817
Elliott S.J.,Wainwright L.A., McDaniel T.K., Jarvis K.G., Deng Y.K., Lai L.C., McNamara B.P., Donnenberg M.S., Kaper J.B. (1998) The complete sequence of the locus of enterocyte effacement (LEE) from enteropathogenic Escherichia coli E2348/69. Mol Microbiol 28: 1–4
Kenny B., DeVinney R., Stein M., Reinscheid D.J., Frey E.A., Finlay B.B. (1997) Enteropathogenic E. coli (EPEC) transfers its receptor for intimate adherence into mammalian cells. Cell 91: 511–520
Hartland E.L., Batchelor M., Delahay R.M., Hale C., Matthews S., Dougan G., Knutton S., Connerton I., Frankel G. (1999) Binding of intimin from enteropathogenic Escherichia coli to Tir and to host cells. Mol Microbiol 32: 151–158
Kenny B., Lai L.C., Finlay B.B., Donnenberg M.S. (1996) EspA, a protein secreted by enteropathogenic Escherichia coli, is required to induce signals in epithelial cells. Mol Microbiol 20: 313–323
Donnenberg M.S., Yu J., Kaper J.B. (1993) A second chromosomal gene necessary for intimate attachment of enteropathogenic Escherichia coli to epithelial cells. J Bacteriol 175: 4670–4680
Lai L.C.,Wainwright L.A., Stone K.D., Donnenberg M.S. (1997) A third secreted protein that is encoded by the enteropathogenic Escherichia coli pathogenicity island is required for transduction of signals and for attaching and effacing activities in host cells. Infect Immun 65: 2211–2217
Crane J.K., McNamara B.P., Donnenberg M.S. (2001) Role of EspF in host cell death induced by enteropathogenic Escherichia coli. Cell Microbiol 3: 197–211
Kenny B., Jepson M. (2000) Targeting of an enteropathogenic Escherichia coli (EPEC) effector protein to host mitochondria. Cell Microbiol 2:579–590
Kenny B. (1999) Phosphorylation of tyrosine 474 of the enteropathogenic Escherichia coli (EPEC) Tir receptor molecule is essential for actin nucleating activity and is preceded by additional host modifications. Mol Microbiol 31: 1229–1241
Batchelor M., Guignot J., Patel A., Cummings N., Cleary J., Knutton S., Holden D.W., Connerton I., Frankel G. (2004) Involvement of the intermediate filament protein cytokeratin-18 in actin pedestal formation during EPEC infection. EMBO Rep 5: 104–110
Goosney D.L., DeVinney R., Finlay B.B. (2001) Recruitment of cytoskeletal and signaling proteins to enteropathogenic and enterohemorrhagic Escherichia coli pedestals. Infect Immun 69: 3315-3322
Rohatgi R., Nollau P., Ho H.Y., Kirschner M.W., Mayer B.J. (2001) Nck and phosphatidylinositol 4,5-bisphosphate synergistically activate actin polymerization through the N-WASP-Arp2/3 pathway. J Biol Chem 276: 26448–26452
Campellone K.G., Giese A., Tipper D.J., Leong J.M. (2002) A tyrosine-phosphorylated 12-amino-acid sequence of enteropathogenic Escherichia coli Tir binds the host adaptor protein Nck and is required for Nck localization to actin pedestals. Mol Microbiol 43:1227–1241
Kalman D.,Weiner O.D., Goosney D.L., Sedat J.W., Finlay B.B., Abo A., Bishop J.M. (1999) Enteropathogenic E. coli acts throughWASP and Arp2/3 complex to form actin pedestals. Nat Cell Biol 1: 389–391
Campellone K.G., Leong J.M. (2003) Tails of two Tirs: actin pedestal formation by enteropathogenic E. coli and enterohemorrhagic E. coli O157:H7. Curr Opin Microbiol 6: 82–90
Lommel S., Benesch S., Rottner K., Franz T.,Wehland J., Kuhn R. (2001) Actin pedestal formation by enteropathogenic Escherichia coli and intracellular motility of Shigella flexneri are abolished in N-WASP-defective cells. EMBO Rep 2: 850–857
Campellone K.G., Robbins D., Leong J.M. (2004b) EspFU is a translocated EHEC effector that interacts with Tir and N-WASP and promotes Nck-independent actin assembly. Dev Cell 7: 217–228
Sambrook J., Russell D. (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Chuang C.H., Hsu S.C., Hsu C.L., Hsu T.C., Syu W.J. (2001) Construction of a tagging system for subcellular localization of proteins encoded by open reading frames. J Biomed Sci 8: 170–175
Campellone K.G., Rankin S., Pawson T., Kirschner M.W., Tipper D.J., Leong J.M. (2004a) Clustering of Nck by a 12-residue Tir phosphopeptide is sufficient to trigger localized actin assembly. J Cell Biol 164: 407–416
Lin H.P., Hsu S.C.,Wu J.C., Sheen I.J., Yan B.S., Syu W.J. (1999) Localization of isoprenylated antigen of hepatitis delta virus by anti-farnesyl antibodies. J Gen Virol 80: 91–96
Hsu S.C., Lin H.P.,Wu J.C., Ko K.L., Sheen I.J., Yan B.S., Chou C.K., SyuW.J. (2000) Characterization of a strain-specific monoclonal antibody to hepatitis delta virus antigen. J Virol Methods 87: 53–62
Knutton S., McConnell M.M., Rowe B., McNeish A.S. (1989) Adhesion and ultrastructural properties of human enterotoxigenic Escherichia coli producing colonization factor antigens III and IV. Infect Immun 57: 3364–3371
Phan-Thanh L., Mahouin F. (1999) A proteomic approach to study the acid response in Listeria monocytogenes. Electrophoresis 20: 2214–2224
Yu S.L., Ko K.L., Chen C.S., Chang Y.C., SyuW.J. (2000) Characterization of the distal tail fiber locus and determination of the receptor for phage AR1, which specifically infects Escherichia coli O157:H7. J Bacteriol 182: 5962–5968
Chiu H.J., LinW.S., Syu W.J., (2003) Type III secretion of EspB in enterohemorrhagic Escherichia coli O157:H7. Arch Microbiol 180: 218–226
Gruenheid S., DeVinney R., Bladt F., Goosney D., Gelkop S., Gish G.D., Pawson T., Finlay B.B. (2001) Enteropathogenic E. coli Tir binds Nck to initiate actin pedestal formation in host cells. Nat Cell Biol 3: 856–859
DeVinney R., Stein M., Reinscheid D., Abe A., Ruschkowski S., Finlay B.B. (1999) Enterohemorrhagic Escherichia coli O157:H7 produces Tir, which is translocated to the host cell membrane but is not tyrosine phosphorylated. Infect Immun 67: 2389–2398
Matsuzawa T., Kuwae A., Yoshida S., Sasakawa C., Abe A. (2004) Enteropathogenic Escherichia coli activates the RhoA signaling pathway via the stimulation of GEF-H1. Embo J 23: 3570–3582
Garmendia J., Phillips A.D., Carlier M.F., Chong Y., Schüller S., Marches O., Dahan S., Oswald E., Shaw R.K., Knutton S., Frankel G. (2004) TccP is an enterohaemorrhagic Escherichia coli O157:H7 type III effector protein that couples Tir to the actin-cytoskeleton. Cell Microbiol 6: 1167–1183
Acknowledgements
This research was supported in parts by Grant 89-B-FA22-2-4 (Program for Promoting Academic Excellence of Universities) from the Department of Education and NSC 92-2320- B-010-048 from the National Science Council, Taiwan, R.O.C.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chuang, CH., Chiu, HJ., Hsu, SC. et al. Comparison of Tir from enterohemorrahgic and enteropathogenic Escherichia coli strains: two homologues with distinct intracellular properties. J Biomed Sci 13, 73–87 (2006). https://doi.org/10.1007/s11373-005-9034-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11373-005-9034-x