Attaching and effacing (A/E) lesion formation by enteropathogenic E. coli on human intestinal mucosa is dependent on non-LEE effectors

Cepeda-Molero, Massiel; Berger, Cédric N.; Walsham, Alistair D. S.; Ellis, Samuel J.; Wemyss‐Holden, Simon A.; Schüller, Stephanie; Frankel, Gad; Fernández, Luis A.

doi:10.1371/journal.ppat.1006706

Cited by 59 publications

(57 citation statements)

References 60 publications

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“…It also agrees with studies that used an equal number of virulence genes to characterise the EHEC pathotype 16,41 . The overall prevalence of EPEC/EHEC (eaeA gene) found in our study is consistent with other studies 18,21,41 . A higher frequency of 42.3% has been reported in India 15 while over 50% has been reported in South African domestic water 6 , 67% in Tennessee 62 and 47.5% found in Iranian children 7 .…”

Section: Discussion Identification and Distribution Of Dec Pathotypessupporting

confidence: 93%

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Antibiotic Resistance Profile and Clonality of E. coli Isolated from Water and Paediatric Stool Samples in the North-West, Province South Africa

Chukwu¹,

Abia²,

Ubomba-Jaswa³

et al. 2019

J Pure Appl Microbiol

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This study investigated the antibacterial resistance profiles of E. coli pathotypes isolated from children under five years and drinking water samples collected from the North West Province of South Africa and ascertained the clonality of the isolates. Two hundred and forty-one E. coli isolates were recovered from stool samples of diarrhoeic and non-diarrhoeic children under five years old, and drinking water, using the Colilert-18 ® Quanti-tray/2000 and Eosin methylene blue agar. The presence of enteropathogenic (eaeA), enterohaemorrhagic (eaeA,stx1, stx2 and flicH7), enteroaggregative (eagg), enteroinvasive (ipaH) and enterotoxigenic (ST and LT) E. coli pathotypes were also investigated using PCR. Antibiotic susceptibility was carried out through the disk diffusion method. The presence of blaCTX-M, blaSHV, blaCMY, and blaDHA genes that code for β-lactamases was investigated using real-time PCR. Similarities between human and water isolates were tested using ERIC-PCR. Overall, EHEC (35.8%), EPEC/EHEC (22%), ETEC (21.6%) and EIEC (20.2%) were detected. The highest antibiotic resistance was detected to Clarithromycin (100%) and Erythromycin (100%) while the lowest resistance was against Gentamicin (0.4%). Also, 100% sensitivity was recorded to imipenem and meropenem. Multi-antibiotic resistance was observed in all the pathotypes, and the ESBL genes were detected in 71.6% of the pathotypes. ERIC-PCR indicated 100% similarities in some water and human samples. Pathogenic E. coli is amongst the diarrhoea-causing agents in the North West Province with EHEC being the most identified pathotype. The clonal relatedness of the human and water isolates suggests that domestic water might be a route of transmission.

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Section: Discussion Identification and Distribution Of Dec Pathotypessupporting

confidence: 93%

“…The eae-encoded intimin proteinis an essential EPEC virulence gene which allows for attaching and effacing (A/E) lesions on the host's intestinal epithelial cell surface 5,18 . Other virulence factors found in tEPEC are the bfpA and perABC genes which encode the bundle-forming pili used for localised attachment to epithelial cells 15,16 .…”

Section: Introductionmentioning

confidence: 99%

Antibiotic Resistance Profile and Clonality of E. coli Isolated from Water and Paediatric Stool Samples in the North-West, Province South Africa

Chukwu¹,

Abia²,

Ubomba-Jaswa³

et al. 2019

J Pure Appl Microbiol

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“…Multiple sequence alignment showed higher nucleotide diversity between the LEE4 and LEE5 operons from BA320 and E2348/69, while the LEE1-3 operons shared higher similarity. Six effector-encoding genes (espG, espZ, espH, map, tir, and espF) are present in the LEE of BA320, of which Tir is the only one absolutely required for AE lesion formation on epithelial cells (35). Analysis of the Tir BA320 sequence identified the NPY motif implicated in the EspFumediated actin polymerization pathway in EHEC O157:H7, while the tyrosine residue Y474, required for Nck-dependent actin polymerization in EPEC O127:H6, was absent ( Fig.…”

Section: Resultsmentioning

confidence: 99%

EspFu-Mediated Actin Assembly Enhances Enteropathogenic Escherichia coli Adherence and Activates Host Cell Inflammatory Signaling Pathways

Martins

Kumar

Abe

et al. 2020

mBio

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The translocation of effectors into the host cell through type 3 secretion systems (T3SS) is a sophisticated strategy employed by pathogenic bacteria to subvert host responses and facilitate colonization. Enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) utilize the Tir and EspFu (also known as TccP) effectors to remodel the host cytoskeleton, culminating in the formation of attaching and effacing (AE) lesions on enterocytes. While some EPEC strains require tyrosine phosphorylation of Tir and recruitment of the host Nck to trigger actin polymerization, EHEC and certain EPEC strains, whose Tir is not phosphorylated, rely on the effector EspFu for efficient actin remodeling. Here, we investigated the role played by Tir-Nck and Tir-EspFu actin polymerization pathways during the infection of epithelial cells, as well as the host transcriptional response to the AE lesion formation induced by EPEC. We found that EspFu-mediated actin assembly promotes bacterial attachment and epithelial colonization more efficiently than Tir-Nck. Moreover, we showed that both actin polymerization mechanisms can activate inflammatory pathways and reverse the anti-inflammatory response induced by EPEC in epithelial cells. However, this activity is remarkably more evident in infections with EspFu-expressing EPEC strains. This study demonstrates the complex interactions between effector-mediated actin remodeling and inflammation. Different strains carry different combinations of these two effectors, highlighting the plasticity of pathogenic E. coli enteric infections. IMPORTANCE EPEC is among the leading causes of diarrheal disease worldwide. The colonization of the gut mucosa by EPEC results in actin pedestal formation at the site of bacterial attachment. These pedestals are referred to as attaching and effacing (AE) lesions. Here, we exploit the different molecular mechanisms used by EPEC to induce AE lesions on epithelial cells, showing that the effector EspFu is associated with increased bacterial attachment and enhanced epithelial colonization compared to the Tir-Nck pathway. Moreover, we also showed that actin pedestal formation can counterbalance the anti-inflammatory activity induced by EPEC, especially when driven by EspFu. Collectively, our findings provide new insights into virulence mechanisms employed by EPEC to colonize epithelial cells, as well as the host response to this enteric pathogen.

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“…The presence of this gene, together with espJ, stx1a or 2a, intimin, and Tir, makes these strains highly pathogenic. Another important illustration of the mosaic of virulence factors found within STEC O26:H11 strains is that two important genes for intestinal colonization (efa1/lifA - a protein with putative glycosyltransferase activity that has an important role in intestinal colonization), and katP- (a catalase-peroxidase, which might help EHEC O157:H7 to colonize host intestines by reducing oxidative stress), were present in both CFSAN027343 and CFSAN027346 (50,51).…”

Section: Discussionmentioning

confidence: 99%

Nanopore sequencing for fast determination of plasmids, phages, virulence markers, and antimicrobial resistance genes in Shiga toxin-producingEscherichia coli

González-Escalona¹,

Allard²,

Brown³

et al. 2019

Preprint

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1Whole genome sequencing can provide essential public health information. However, it is now 2 known that widely used short-read methods have the potential to miss some randomly-3 distributed segments of genomes. This can prevent phages, plasmids, and virulence factors 4 from being detected or properly identified. Here, we compared assemblies of three complete 5 STEC O26:H11 genomes from two different sequence types (ST21 and 29), each acquired 6 using the MiSeq-Nextera XT, MinION nanopore-based sequencing, and Pacific Biosciences 7 (PacBio) sequencing. Each closed genome consisted of a single chromosome, approximately 8 5.7 Mb for CFSAN027343, 5.6 Mb for CFSAN027346, and 5.4 MB for CFSAN027350. However, 9 short-read WGS using MiSeq-Nextera failed to identify some virulence genes in plasmids and 10 on the chromosome, both of which were detected using the long-read platforms. Results from 11 long-read MinION and PacBio allowed us to identify differences in plasmid content: a single 88 12 kb plasmid in CFSAN027343; a 157kb plasmid in CFSAN027350; and two plasmids in 13 CFSAN027346 (one 95 Kb, one 72 Kb). These data enabled rapid characterization of the 14 virulome, detection of antimicrobial genes, and composition/location of Stx phages. Taken 15 together, positive correlations between the two long-read methods for determining plasmids, 16 virulome, antimicrobial resistance genes, and phage composition support MinION sequencing 17 as one accurate and economical option for closing STEC genomes and identifying specific 18 virulence markers. 12are very complex, containing many virulence genes, insertion sequences, phages, and 13 plasmids; consequently, strains of the same lineage can possess significantly different content 14 (7,9,35), (33,(36)(37)(38). Missing the presence of some of these elements during an investigation can 15 have large impacts on human health (e.g. hlyA gene in EHECs). Thus, it really matters that we 16 have reproducible WGS systems for fully capturing and sequencing these elements. 18Long-read sequencing platforms afford one solution to this challenge. Some systems such as 19 Pacific Biosciences (PacBio) Sequencers RSII or Sequel (https://www.pacb.com/products-and-20 services/pacbio-systems/), use single-molecule real-time (SMRT) sequencing technology that 21 allow for real-time observation of DNA synthesis through zero-mode waveguides (ZMWs) and 22 phospho-linked nucleotides (11,12). While comprehensive in their ability to capture entire 23 genomes, extraneous elements included, these systems often require significant investments in 24 machinery, space, and laboratory expertise, all of which may be obstacles to routine use. These 25 systems also require significant quantities of DNA (i.e., 5 µg), require a more substantial preparatory time (i.e., 8 hr DNA sequencing library protocols, and produce average read lengths 1 of about 11Kb, although reads of greater than 50 kb were possible in our laboratory at the time 2 of this study. 4Alternative sequencing platforms based on nanopore technology ma...

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Attaching and effacing (A/E) lesion formation by enteropathogenic E. coli on human intestinal mucosa is dependent on non-LEE effectors

Cited by 59 publications

References 60 publications

Antibiotic Resistance Profile and Clonality of E. coli Isolated from Water and Paediatric Stool Samples in the North-West, Province South Africa

Antibiotic Resistance Profile and Clonality of E. coli Isolated from Water and Paediatric Stool Samples in the North-West, Province South Africa

EspFu-Mediated Actin Assembly Enhances Enteropathogenic Escherichia coli Adherence and Activates Host Cell Inflammatory Signaling Pathways

Nanopore sequencing for fast determination of plasmids, phages, virulence markers, and antimicrobial resistance genes in Shiga toxin-producingEscherichia coli

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