Functional analysis of <scp><i>E</i></scp><i>scherichia coli</i><scp>Y</scp>ad fimbriae reveals their potential role in environmental persistence

Larsonneur, Fanny; Martín, Fernando A.; Mallet, Adeline; Martínez-Gil, Marta; Semetey, Vincent; Ghigo, Jean‐Marc; Beloin, Christophe

doi:10.1111/1462-2920.13559

Cited by 27 publications

(33 citation statements)

References 85 publications

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“…Here we visualized, to our knowledge for the first time, surface-expressed Yeh fimbriae of E. coli. The Yad fimbriae of E. coli O157:H7 Sakai showed a similar appearance compared to Yad fimbriae encoded by E. coli K-12 (Korea et al, 2010;Larsonneur et al, 2016). Expression of Curli fimbriae and type 4 pili revealed diffuse adhesive structures on the bacterial cell surface.…”

Section: Discussionmentioning

confidence: 87%

“…Genes for yad fimbriae are present in several serotypes of E. coli, ranging from non-pathogenic E. coli K-12 to pathogenic serotypes UPEC, APEC, and EHEC (Wurpel et al, 2013). Expression of Yad fimbriae of E. coli K-12 was observed at ambient temperatures (RT), under anaerobic conditions, as well as during biofilm formation (Larsonneur et al, 2016). In contrast, Yad fimbriae encoded by APEC strain SCI-07 causing chicken infections, are expressed at higher temperatures of 41 • C (Verma et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

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Expression and Functional Characterization of Various Chaperon-Usher Fimbriae, Curli Fimbriae, and Type 4 Pili of Enterohemorrhagic Escherichia coli O157:H7 Sakai

Elpers

Hensel

2020

Front. Microbiol.

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Enterohemorrhagic Escherichia coli (EHEC) is a highly pathogenic strain leading to hemorrhagic colitis and to the hemolytic-uremic syndrome (HUS) in humans. The mechanisms by which pathogenic E. coli infect and colonize humans leading to the typical disease pattern are in focus of many investigations. The adhesion of EHEC to epithelial cells by the coordinated translocation of receptor Tir and surface expression of corresponding adhesin intimin is a key event in host-pathogen-interaction. However, less is known about other adhesins encoded by EHEC, especially about the complex set of fimbrial adhesins varying among various serotypes. Here, we investigate EHEC serotype O157:H7 strain Sakai possessing at least 16 putative fimbrial gene clusters. Using a synthetic heterologous expression system in a non-pathogenic E. coli strain, a subset of 6 gene clusters for fimbrial adhesins was analyzed. We were able to visualize surface expression of two γ1 class fimbriae (Fim and Ycb), two γ4 class fimbriae (Yad and Yeh), and two fimbrial adhesins which are assembled by the nucleation/precipitation pathway (Curli fimbriae), and by a type 2 secretion system (type 4 pili). Further, we elucidated the impact of these fimbrial adhesins in adhesion to various epithelial cells lines (HeLa, MDCK, and CaCo2), and the contribution on biofilm formation. We demonstrate the ultrastructure of Fim fimbriae and Yad fimbriae of EHEC Sakai, and Yeh fimbriae of E. coli in general. The involvement of Fim fimbriae of EHEC Sakai to adhesion to various epithelial cell lines, and contribution to biofilm formation is reported here. Our approach provides first ultrastructural and functional data for novel EHEC adhesins, and enables further understanding of the involvement of fimbrial adhesins in pathogenesis of EHEC Sakai.

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Section: Discussionmentioning

confidence: 87%

Section: Discussionmentioning

confidence: 99%

Expression and Functional Characterization of Various Chaperon-Usher Fimbriae, Curli Fimbriae, and Type 4 Pili of Enterohemorrhagic Escherichia coli O157:H7 Sakai

Elpers

Hensel

2020

Front. Microbiol.

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“…Cultures grown overnight in BHIS medium were washed and resuspended in 1ϫ PBS. "Shaving" of surface pili was performed as previously described (45). Briefly, 100 ml of the culture was grown in BHIS medium for 48 h at 37°C without agitation under anaerobic conditions.…”

Section: Methodsmentioning

confidence: 99%

A Putative Type V Pilus Contributes to Bacteroides thetaiotaomicron Biofilm Formation Capacity

et al. 2019

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Bacteroides thetaiotaomicron is a prominent anaerobic member of the healthy human gut microbiota. While the majority of functional studies on B. thetaiotaomicron addressed its impact on the immune system and the utilization of diet polysaccharides, B. thetaiotaomicron biofilm capacity and its contribution to intestinal colonization are still poorly characterized. We tested the natural adhesion of 34 B. thetaiotaomicron isolates and showed that although biofilm capacity is widespread among B. thetaiotaomicron strains, this phenotype is masked or repressed in the widely used reference strain VPI 5482. Using transposon mutagenesis followed by a biofilm positive-selection procedure, we identified VPI 5482 mutants with increased biofilm capacity corresponding to an alteration in the C-terminal region of BT3147, encoded by the BT3148-BT3147 locus, which displays homology with Mfa-like type V pili found in many Bacteroidetes. We show that BT3147 is exposed on the B. thetaiotaomicron surface and that BT3147-dependent adhesion also requires BT3148, suggesting that BT3148 and BT3147 correspond to the anchor and stalk subunits of a new type V pilus involved in B. thetaiotaomicron adhesion. This study therefore introduces B. thetaiotaomicron as a model to study proteinaceous adhesins and biofilm-related phenotypes in this important intestinal symbiont. IMPORTANCE Although the gut anaerobe Bacteroides thetaiotaomicron is a prominent member of the healthy human gut microbiota, little is known about its capacity to adhere to surfaces and form biofilms. Here, we identify that alteration of a surface-exposed protein corresponding to a type of pili found in many Bacteroidetes increases B. thetaiotaomicron biofilm formation. This study lays the ground for establishing this bacterium as a model organism for in vitro and in vivo studies of biofilm-related phenotypes in gut anaerobes.

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“…Adherence is a key step in early interactions with host tissue and STEC fimbrial adhesins that mediate specific binding to plant cell wall components include E. coli common pilus (ECP) and Yad fimbriae [14, 43] and non-specific interactions via flagella [15]. Potential candidates enriched in the screen may be involved in non-adherence functions, such as response to PAMP perception by the host, Nle effectors since NleA is known to play a role in disrupting secretory pathways [44, 45] or modulating host cytoskeleton (EspO1-2) [46] in animal hosts.…”

Section: Resultsmentioning

confidence: 99%

A high-throughput genomic screen identifies a role for the plasmid-borne Type II secretion system ofEscherichiacoli O157:H7 (Sakai) in plant-microbe interactions

Holmes

Pritchard

Hedley

et al. 2020

Preprint

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Food-borne illness arising from Shiga-toxigenic Escherichia coli (STEC) is often linked to consumption of fruit and vegetables as the bacteria have the ability to interact with plants and use them as alternative or secondary hosts. The initial stages of the interaction involve chemotaxis, attachment and potentially, responding to the early stages of microbe perception by the plant host. We used a high-throughput positive-selection approach to identify early interaction factors of E. coli O157:H7 isolate Sakai to spinach. A bacterial artificial chromosome (BAC) clone library was quantified by microarray hybridisation, and gene loci enrichment measured using a Bayesian hierarchical model. The screen of four successive rounds of short-term (2 hour) interaction with spinach roots produced in 115 CDS credible candidates, comprising seven contiguous genomic regions. Two candidate regions were selected for functional assessment: a chaperone-usher fimbrial gene cluster (loc6) and the pO157 plasmid-encoded type two secretion system (T2SS). Interaction of bacteria with spinach tissue was reduced in the absence of the pO157 plasmid, which was appeared to involve the T2SS EtpD secretin protein, whereas loss of loc6 did not impact interactions. The T2SS genes, etpD and etpC, were expressed at a plant-relevant temperature of 18 °C, and etpD expressed in planta by E. coli Sakai on spinach plants. Thus, a whole genome screening approach using a combination of computational modelling and functional assays has identified a novel function for STEC T2SS in interactions with plant tissue.

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Functional analysis of Escherichia coliYad fimbriae reveals their potential role in environmental persistence

Cited by 27 publications

References 85 publications

Expression and Functional Characterization of Various Chaperon-Usher Fimbriae, Curli Fimbriae, and Type 4 Pili of Enterohemorrhagic Escherichia coli O157:H7 Sakai

Expression and Functional Characterization of Various Chaperon-Usher Fimbriae, Curli Fimbriae, and Type 4 Pili of Enterohemorrhagic Escherichia coli O157:H7 Sakai

A Putative Type V Pilus Contributes to Bacteroides thetaiotaomicron Biofilm Formation Capacity

A high-throughput genomic screen identifies a role for the plasmid-borne Type II secretion system ofEscherichiacoli O157:H7 (Sakai) in plant-microbe interactions

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