Cytokeratin 18 interacts with the enteropathogenic Escherichia coli secreted protein F (EspF) and is redistributed after infection

Virinchipuram Viswanathan, Sandra Lukic, Athanasia Koutsouris, Richard Miao, Michelle M. Muza, Gail Hecht

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Enteropathogenic Escherichia coli (EPEC) pathogenesis requires the delivery of effector proteins into host cytosol by a type III secretion system. The effector protein EspF, while critical for disruption of epithelial barrier function through alteration of tight junctions, is not required for bacterial viability or attachment. Yeast two-hybrid analyses revealed host intermediate filament (IF) protein cytokeratin 18 (CK18) as an interacting partner of EspF. This was confirmed by co-immunoprecipitation of extracts from EPEC-infected epithelial cells. EPEC infection increased detergent-soluble CK18 amounts without significantly altering CK18 expression. The adaptor protein 14-3-3 binds to CK18 and modulates its solubility. EPEC infection promoted CK18/14-3-3 interactions, corresponding to the increase of CK18 in the soluble fractions. 14-3-3 also co-immunoprecipitated with EspF, suggesting that CK18, 14-3-3 and EspF may form a complex in infected cells. The 14-3-3ζ isoform was co-immunoprecipitated with CK18, suggesting the involvement of specific signalling pathways. Immunofluorescence studies revealed a dramatic alteration in the architecture of the IF network in EPEC-infected epithelial cells. IF fragmentation, evident at 2 h post infection, progressed to a collapse of this network at later time points. The secretion mutant (ΔescN) failed to alter CK18 solubility and IF morphology, while deletion of espF partially impaired the ability of EPEC to induce CK18 modifications. These results suggest that modifications in 14-3-3 interactions and IF network, modulated by type III secreted proteins, may be an important step in EPEC pathogenesis.

Original languageEnglish (US)
Pages (from-to)987-997
Number of pages11
JournalCellular Microbiology
Volume6
Issue number10
DOIs
StatePublished - Oct 2004
Externally publishedYes

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Keratin-18
Enteropathogenic Escherichia coli
Escherichia coli Proteins
Escherichia coli
Infection
Intermediate Filaments
Proteins
Keratin-14
Escherichia coli Infections
Solubility
Epithelial Cells
Microbial Viability
14-3-3 Proteins
Intermediate Filament Proteins
Tight Junctions
Immunoprecipitation
Detergents
Cytosol
Yeast
Fluorescent Antibody Technique

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Microbiology

Cite this

Cytokeratin 18 interacts with the enteropathogenic Escherichia coli secreted protein F (EspF) and is redistributed after infection. / Viswanathan, Virinchipuram; Lukic, Sandra; Koutsouris, Athanasia; Miao, Richard; Muza, Michelle M.; Hecht, Gail.

In: Cellular Microbiology, Vol. 6, No. 10, 10.2004, p. 987-997.

Research output: Contribution to journalArticle

Viswanathan, Virinchipuram ; Lukic, Sandra ; Koutsouris, Athanasia ; Miao, Richard ; Muza, Michelle M. ; Hecht, Gail. / Cytokeratin 18 interacts with the enteropathogenic Escherichia coli secreted protein F (EspF) and is redistributed after infection. In: Cellular Microbiology. 2004 ; Vol. 6, No. 10. pp. 987-997.
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