Involvement of the intermediate filament protein cytokeratin‐18 in actin pedestal formation during EPEC infection

Batchelor, Miranda; Guignot, Julie; Patel, Amit R.; Cummings, Nicola J.; Cleary, Johanna; Knutton, Stuart; Holden, David W.; Connerton, Ian F.; Frankel, Gad

doi:10.1038/sj.embor.7400087

Cited by 22 publications

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“…Only those extracts prepared from wild‐type EPEC contained Tir. Cytokeratin 18 was included as a control because it has been shown to independently bind other 14‐3‐3 isoforms and Tir (Ku et al ., 1998; Batchelor et al ., 2004). A mouse monoclonal antibody directed against Tir was used to immunoprecipitate Tir and associated proteins in conjunction with immobilized protein A.…”

Section: Resultsmentioning

confidence: 99%

“…Although over the course of the experiment the disrupted HeLa cells did not demonstrate any observable growth defect, they were clearly deficient in their ability to mobilize actin for pedestal formation in response to EPEC. Recently it has been reported that EPEC infection may promote 14‐3‐3ζ interaction with cytokeratin 18 (Viswanathan et al ., 2004), an intermediate filament protein that also binds Tir (Batchelor et al ., 2004). The formation of functional 14‐3‐3 heterodimers in the regulation of the cytoskeletal remodelling required to form pedestal structure would seem a real possibility.…”

Section: Discussionmentioning

confidence: 99%

“…Enteropathogenic Escherichia coli (EPEC) belong to a group of pathogens that cause distinctive lesions in the host intestine, termed attaching‐effacing (A/E) lesions (Ulshen and Rollo, 1980; Moon et al ., 1983; Phillips et al ., 2000). The A/E lesion is characterized by localized destruction (effacement) of brush border microvilli, intimate attachment of the bacillus to the host cell plasma membrane and the formation of an underlying pedestal‐like structure in the host cell consisting of polymerized actin, ezrin, talin and myosin (Goosney et al ., 2001), as well as WASP (Wiskott–Aldrich syndrome family of proteins), Arp2/3 complex (Kalman et al ., 1999) and cytokeratins 8/18 (Batchelor et al ., 2004). All the bacterial genes required for A/E lesion formation are present in a large pathogenicity island, termed the locus of enterocyte effacement or LEE (McDaniel and Kaper, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Host protein interactions with enteropathogenic Escherichia coli (EPEC): 14-3-3tau binds Tir and has a role in EPEC-induced actin polymerization

et al. 2006

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SummaryEnteropathogenic Escherichia coli (EPEC) cause infantile diarrhoea and are characterized by their ability to produce attaching and effacing lesions on the surface of intestinal epithelial cells. EPEC employ a filamentous type III secretion system to deliver effector molecules that subvert mammalian cell function to generate actin-and cytokeratin-rich pedestals beneath adherent bacteria. Tir is a major effector protein that is delivered to the plasma membrane of the eukaryotic cell where it acts as the receptor for the bacterial adhesin intimin. Host cell proteins that are recruited to the site of intimate attachment include focal adhesion and cytoskeletal proteins that contribute to pedestal formation. We have used Tir as bait in a yeast two-hybrid screen to identify the protein 14-3-3 t t t t as a binding partner. 14-3-3 proteins are a family of adaptor proteins that modulate protein function in all eukaryotic cells. Here we demonstrate that the tau isoform (also known as theta) of 14-3-3 can bind specifically to Tir in a phosphorylation-independent manner, and that the interaction occurs during the infection process by co-immunoprecipitation of the partners from infected HeLa cell extracts. 14-3-3 t t t t is recruited to the site of the pedestal (3 h after infection) and can decorate attached EPEC in the later stages of the infection process (5-7 h). Pedestal formation can be impaired by depletion of cellular 14-3-3 t t t t using small interfering RNAs. This study indicates a direct functional role for the 14-3-3 t t t t :Tir interaction and is the first to demonstrate the association of a host protein with the surface of EPEC.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Host protein interactions with enteropathogenic Escherichia coli (EPEC): 14-3-3tau binds Tir and has a role in EPEC-induced actin polymerization

et al. 2006

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“…It is widely accepted that the cytoskeleton and intercellular junctions are targeted by A/E pathogens (Spitz et al, 1995; Guttman et al, 2006a). Apart from the obvious alterations to the actin filament component of the cytoskeleton in pedestals, microtubules and intermediate filaments are also modified (Batchelor et al, 2004; Viswanathan et al, 2004b; Hardwidge et al, 2005; Tomson et al, 2005). Two studies have investigated the intermediate filament cytoskeleton and its modification during EPEC infection.…”

mentioning

confidence: 99%

“…Two studies have investigated the intermediate filament cytoskeleton and its modification during EPEC infection. Batchelor et al (2004) demonstrated that the intermediate filament protein cytokeratin‐18 is found in EPEC pedestals. Viswanathan et al (2004b) assessed the overall effects on cytokeratin‐18 during EPEC infection of T84 cells.…”

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Desmosomes are unaltered during infections by attaching and effacing pathogens

Guttman

Kazemi

Lin

et al. 2007

The Anatomical Record

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The human attaching and effacing (A/E) intestinal pathogens enterohemorrhagic Escherichia coli (EHEC), enteropathogenic E. coli (EPEC), and the murine A/E pathogen Citrobacter rodentium cause serious diarrhea in their hosts. These bacteria alter numerous host cell components, including organelles, the host cell cytoskeleton, and tight junctions during the infectious process. One of the proteins that contribute to the intermediate filament network in host cells, cytokeratin-18, is extensively altered during EPEC infections.Based on this, we tested the hypothesis that desmosomes, the only intercellular junctions that interact with intermediate filaments, are also influenced by A/E pathogen infections. We found that the desmosomal transmembrane proteins desmoglein and desmocollin, as well as the desmosome plaque protein desmoplakin, all remain unchanged during EPEC infection in vitro. This evidence is corroborated by the unaltered localization of desmoglein and desmoplakin in vivo in mice infected with C. rodentium for 7 days. Electron microscopic analysis of 7-day C. rodentium-infected murine colonocytes also show no observable differences in the desmosomes when compared to uninfected controls. Our data suggest that, unlike tight junctions, the desmosome protein levels and localization, as well as desmosome morphology, are unaltered during A/E pathogenesis.

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Distribution of CD147 During EnteropathogenicEscherichia coliandSalmonella entericaSerovar Typhimurium Infections

Dhanda

Guttman

2019

The Anatomical Record

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Enteropathogenic Escherichia coli (EPEC) and Salmonella enterica serovar Typhimurium (S. Typhimurium) are highly infectious gastrointestinal human pathogens. These microbes inject bacterial‐derived effector proteins directly into the host cell cytosol as part of their disease processes. A common host subcellular target of these pathogens is the actin cytoskeleton, which is commandeered by the bacteria and is used during their attachment onto (EPEC) or invasion into (S. Typhimurium) the host cells. We previously demonstrated that the host enzyme cyclophilin A (CypA) is recruited to the actin‐rich regions of EPEC pedestals and S. Typhimurium membrane ruffles. To further expand the growing catalogue of host proteins usurped by actin‐hijacking bacteria, we examined the host plasma membrane protein and cognate receptor of CypA, CD147, during EPEC and S. Typhimurium infections. Here, we show that CD147 is enriched at the basolateral regions of pedestals but, unlike CypA, it is absent from their actin‐rich core. We show that the CD147 recruitment to these areas requires EPEC pedestal formation and not solely bacteria‐host cell contact. Additionally, we demonstrate that the depletion of CD147 by siRNA does not alter the formation of pedestals. Finally, we show that CD147 is also a component of actin‐rich membrane ruffles generated during S. Typhimurium invasion of host cells. Collectively, our findings establish CD147 as another host component present at dynamic actin‐rich structures formed during bacterial infections. Anat Rec, 302:2224–2232, 2019. © 2019 American Association for Anatomy

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Involvement of the intermediate filament protein cytokeratin‐18 in actin pedestal formation during EPEC infection

Abstract: In the January 2004 issue, we published an error in Fig 4. The CK18 and CK8 labels that appeared in the two top left panels were added mistakenly, also the label in row 4 column 1 was incomplete. The figure should have been published as shown below.

Cited by 22 publications

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Host protein interactions with enteropathogenic Escherichia coli (EPEC): 14-3-3tau binds Tir and has a role in EPEC-induced actin polymerization

Host protein interactions with enteropathogenic Escherichia coli (EPEC): 14-3-3tau binds Tir and has a role in EPEC-induced actin polymerization

Desmosomes are unaltered during infections by attaching and effacing pathogens

Distribution of CD147 During EnteropathogenicEscherichia coliandSalmonella entericaSerovar Typhimurium Infections

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