Contribution of Efa1/LifA to the adherence of enteropathogenic Escherichia coli to epithelial cells

Badea, Luminita; Doughty, Stephen W.; Nicholls, Larissa; Sloan, Joan; Robins-Browne, Roy M.; Hartland, Elizabeth L.

doi:10.1016/s0882-4010(03)00026-3

Cited by 66 publications

(61 citation statements)

References 30 publications

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“…Our results that ToxB is a type III effector may also explain the paradox that LifA, an important virulence factor in C. rodentium and EHEC O5 and O111 strains (38 -40), was truncated into a much shorter form (Z4332) in the highly virulent, outbreak-causing EHEC O157:H7 strains (22,42), whereas the entire ORF of LifA/Efa1 is present in all other A/E pathogens surveyed (50). We have shown that Z4332 is an effector, but Z4332 does not contain either the glycosyltransferase motif DXD or the other functional domains of LifA (Fig.…”

Section: Discussionmentioning

confidence: 64%

“…The nearly identical LifA and Efa1 are encoded only by the eae (therefore LEE)-positive A/E pathogens, and their homologs, including the truncated Z4332 and Z4333 in EHEC O157:H7 strains, are present in almost all of the A/E pathogens analyzed (37,38,50). ToxB is encoded on the pO157 plasmid in EHEC O157:H7 strains (41).…”

Section: Discussionmentioning

confidence: 99%

“…However, LifA/Efa1 proteins do not possess these C-terminal translocation and receptorbinding domains of the clostridial toxins. Efa1 was originally identified as an exported protein, at least to the bacterial periplasm, by TnphoA mutagenesis (38), and there was some evidence by immunofluorescence that Efa1 is associated with the membranes (50). However, LifA/Efa1/ToxB do not have a typical signal sequence for exported proteins at their N termini, and how they gain entry and act within target cells is still unresolved.…”

Section: Discussionmentioning

confidence: 99%

“…3A). We were able to verify T3SS-dependent secretion for at least 13 50 , C_2649, Spy, ChuT, FkpA, YehE, YgiW, YebF, YdgH, YbgF, C_4114, ModA, YcdO, ZnuA, FepB, and HdeB were cloned into the vector pCX341 to generate N-terminal fusions to TEM-1. These constructs were introduced into EPEC WT, ⌬escN, and ⌬sepD strains and assayed for type III secretion of the TEM-1 fusion proteins.…”

Section: Experimental Strategy For Silac Analysis Of Epec Type IIImentioning

confidence: 99%

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Quantitative Proteomic Analysis of Type III Secretome of Enteropathogenic Escherichia coli Reveals an Expanded Effector Repertoire for Attaching/Effacing Bacterial Pathogens

Deng

Hoog

et al. 2012

Molecular & Cellular Proteomics

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Type III secretion systems are central to the pathogenesis and virulence of many important Gram-negative bacterial pathogens, and elucidation of the secretion mechanism and identification of the secreted substrates are critical to our understanding of their pathogenic mechanisms and developing potential therapeutics. Stable isotope labeling with amino acids in cell culture-based mass spectrometry is a quantitative and highly sensitive proteomics tool that we have previously used to successfully analyze the type III secretomes of Citrobacter rodentium and Salmonella enterica serovar Typhimurium. In this report, stable isotope labeling with amino acids in cell culture was used to analyze the type III secretome of enteropathogenic Escherichia coli (EPEC), an important human pathogen, which, together with enterohemorrhagic E. coli and C. rodentium, represents the family of attaching and effacing bacterial pathogens. We not only confirmed all 25 known EPEC type III-secreted proteins and effectors previously identified by conventional molecular and bioinformatical techniques but also identified several new type III-secreted proteins, including two novel effectors, C_0814/NleJ and LifA, that were shown to be translocated into host cells. LifA is a known virulence factor believed to act as a toxin as well as an adhesin, but its mechanism of secretion and function is not understood. With a predicted molecular mass of 366 kDa, LifA is the largest type III effector identified thus far in any pathogen. We further demonstrated that Efa1, ToxB, and Z4332 (homologs of LifA in enterohemorrhagic E. coli) are also type III effectors. This study has comprehensively characterized the type III secretome of EPEC, expanded the repertoire of type III-secreted effectors for the attaching and effacing pathogens, and provided new insights into the mode of function for LifA/Efa1/ToxB/Z4332, an important family of virulence factors. Molecular & Cellular

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Section: Discussionmentioning

confidence: 64%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Experimental Strategy For Silac Analysis Of Epec Type IIImentioning

confidence: 99%

See 2 more Smart Citations

Quantitative Proteomic Analysis of Type III Secretome of Enteropathogenic Escherichia coli Reveals an Expanded Effector Repertoire for Attaching/Effacing Bacterial Pathogens

Deng

Hoog

et al. 2012

Molecular & Cellular Proteomics

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“…However, adherence of E. coli strains is also mediated by nonpilus factors. The EHEC factor for adherence (Efa-1) is a nonpilus adhesin that has been implicated in both in vitro adherence and in vivo colonization of the bovine intestinal epithelium (33,446,572). In addition to being a conduit for transport of secreted effector proteins, the EspA filament of the LEE-encoded T3SS has also been implicated in adherence of EPEC strains (103).…”

Section: Adherence To Gastrointestinal Epitheliummentioning

confidence: 99%

Change Is Good: Variations in Common Biological Mechanisms in the Epsilonproteobacterial GeneraCampylobacterandHelicobacter

Gilbreath

Cody

Merrell

et al. 2011

Microbiol Mol Biol Rev

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SUMMARY Microbial evolution and subsequent species diversification enable bacterial organisms to perform common biological processes by a variety of means. The epsilonproteobacteria are a diverse class of prokaryotes that thrive in diverse habitats. Many of these environmental niches are labeled as extreme, whereas other niches include various sites within human, animal, and insect hosts. Some epsilonproteobacteria, such as Campylobacter jejuni and Helicobacter pylori , are common pathogens of humans that inhabit specific regions of the gastrointestinal tract. As such, the biological processes of pathogenic Campylobacter and Helicobacter spp. are often modeled after those of common enteric pathogens such as Salmonella spp. and Escherichia coli . While many exquisite biological mechanisms involving biochemical processes, genetic regulatory pathways, and pathogenesis of disease have been elucidated from studies of Salmonella spp. and E. coli , these paradigms often do not apply to the same processes in the epsilonproteobacteria. Instead, these bacteria often display extensive variation in common biological mechanisms relative to those of other prototypical bacteria. In this review, five biological processes of commonly studied model bacterial species are compared to those of the epsilonproteobacteria C. jejuni and H. pylori . Distinct differences in the processes of flagellar biosynthesis, DNA uptake and recombination, iron homeostasis, interaction with epithelial cells, and protein glycosylation are highlighted. Collectively, these studies support a broader view of the vast repertoire of biological mechanisms employed by bacteria and suggest that future studies of the epsilonproteobacteria will continue to provide novel and interesting information regarding prokaryotic cellular biology.

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Interaction of Enterohemorrhagic Escherichia coli (EHEC) with Mammalian Cells: Cell Adhesion, Type III Secretion, and Actin Pedestal Formation

2007

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Infection by the food‐borne pathogen enterohemorragic Escherichia coli (EHEC) can lead to diarrhea, hemorrhagic colitis, and, in the most serious cases, renal failure. A critical step in colonization is the unusual interaction between EHEC and the intestinal epithelium. EHEC is able to adhere to mammalian cells, and then, by injecting bacterial proteins, or “effectors,” into the host cell via a type III secretion system, induces the formation of attaching and effacing (AE) lesions characterized by the accumulation of filamentous actin directly beneath bound bacteria. This unit describes methods to evaluate the ability of EHEC to adhere to cultured mammalian cells, to secrete type III effectors, and to promote the formation of actin “pedestals.” These methods can not only be used to evaluate the contribution of specific EHEC gene products to adherence, type III secretion, and mammalian cell signaling, but also facilitate the analysis of the eukaryotic pathways controlling fundamental cellular processes such as actin assembly.

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Contribution of Efa1/LifA to the adherence of enteropathogenic Escherichia coli to epithelial cells

Cited by 66 publications

References 30 publications

Quantitative Proteomic Analysis of Type III Secretome of Enteropathogenic Escherichia coli Reveals an Expanded Effector Repertoire for Attaching/Effacing Bacterial Pathogens

Quantitative Proteomic Analysis of Type III Secretome of Enteropathogenic Escherichia coli Reveals an Expanded Effector Repertoire for Attaching/Effacing Bacterial Pathogens

Change Is Good: Variations in Common Biological Mechanisms in the Epsilonproteobacterial GeneraCampylobacterandHelicobacter

Interaction of Enterohemorrhagic Escherichia coli (EHEC) with Mammalian Cells: Cell Adhesion, Type III Secretion, and Actin Pedestal Formation

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