Dataset Information

The role of inverse autotransporter FdeC in Avian Pathogenic Escherichia coli adhesion

ABSTRACT: Avian Pathogenic Escherichia coli (APEC) poses a significant threat to the global poultry industry and has also zoonotic potential, meaning it could infect humans. One critical factor in APEC infection is its ability to adhere to host tissues, a process mediated by a protein called FdeC among others. This protein is present in pathogenic E. coli strains but is often nonfunctional in nonpathogenic ones. Our study focused on understanding the role of FdeC in APEC's ability to bind to chicken intestinal cells. Through an extensive screen of 2000 transposon mutants from the APEC IMT5155 strain, we discovered that a specific mutation in the fdeC gene led to enhanced adhesion. However, a direct deletion of the fdeC gene did not produce the same effect under standard conditions, which led us to investigate the environmental factors that might regulate FdeC expression. We found that FdeC's expression is highly dependent on the environmental conditions such as temperature and pH. Using optimized conditions for FdeC expression, we observed significantly higher adhesion and motility levels in the fdeC-deleted IMT5155 strain compared to the wild type. To understand the mechanisms behind this, we employed mass spectrometry and discovered that the absence of FdeC led to changes in ion transport and downregulated a protein called YbjN, which usually inhibits bacterial movement. Collectively, our findings suggest that FdeC has host-dependent expression and contributes to enhancement of bacterial fitness by modulating motility during colonization.

INSTRUMENT(S): Orbitrap Exploris 480

ORGANISM(S): Escherichia Coli

SUBMITTER: Fernando Garcia

LAB HEAD: Fernando Garcia Martinez

PROVIDER: PXD045075 | Pride | 2024-06-16

REPOSITORIES: Pride

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Publications

FdeC expression regulates motility and adhesion of the avian pathogenic Escherichia coli strain IMT5155.

Aleksandrowicz Adrianna A Kjærup Rikke Brødsgaard RB Grzymajło Krzysztof K Martinez Fernando Garcia FG Muñoz Javier J Borowska Dominika D Sives Samantha S Vervelde Lonneke L Dalgaard Tina Sørensen TS Kingsley Robert A RA Kolenda Rafał R

Veterinary research 20240531 1

Adaptation of avian pathogenic E. coli (APEC) to changing host environments including virulence factors expression is vital for disease progression. FdeC is an autotransporter adhesin that plays a role in uropathogenic Escherichia coli (UPEC) adhesion to epithelial cells. Expression of fdeC is known to be regulated by environmental conditions in UPEC and Shiga toxin-producing E. coli (STEC). The observation in a previous study that an APEC strain IMT5155 in which the fdeC gene was disrupted by a ...[more]

PMID: 38822378

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Project description:Enterohemorrhagic Escherichia coli (EHEC), including serotype O157:H7, cause severe food-borne illness. On route to the human colon, they encounter and resist, numerous anti-microbial ingestion stresses. We hypothesize that these stresses cue EHEC to alter virulence properties. This study investigated the impact of bile salts on virulence properties and examined the genetic basis of the phenotypes. Established assays were used to examine adhesion to human epithelial cells, motility, verotoxin (VT) production and antimicrobial resistance with/without bile salt stress. Bacteria treated for 90 minute in DMEM plus 0.15% (w/v) bile salt mix demonstrated significantly enhanced adhesion to epithelial cells and resistance to several antibiotics but did not increase motility or VT production. To determine the genetic basis of these phenotypes a microarray experiment was conducted. EHEC strain 86-24, in mid-log phase of growth, were grown in DMEM pH 7.4 (control), or DMEM plus bile salt mix (0.15% w/v), for 90 minutes, statically at 37˚C, 5% CO2 prior to harvesting RNA for the microarray study. Four biological replicates were produced for each treatment. Microarray and gene expression analysis (semi-quantitative RT-PCR and beta-galactosidase reporter assays) of bile salt-treated EHEC revealed significant up-regulation of genes for lipid A modification, fimbriae, an efflux pump, and a two-component regulatory system relative to the bacteria grown in DMEM alone. This work points to several mechanisms that EHEC employs to resist the stresses of the human small intestine, notably efflux, antimicrobial resistance, and outer membrane alterations. Bile salts enhanced the virulence-related properties of increased adhesion and resistance to antimicrobials but not VT production or motility. This research contributes to our understanding of how EHEC senses and responds to host environmental signals and the mechanisms this pathogen uses to successfully colonize and infect the human host. Bacteria were grown in LB broth overnight with shaking, then subcultured into DMEM and grown statically at 37˚C, 5%CO2 to mid-log phase. Bacteria were then subjected to one of two 90 minute treatments: 1) Control: DMEM pH 7.4, or 2) Bile Salt Stress: DMEM pH 7.4 plus 0.15%, grown statically at 37˚C, 5%CO2.

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Dataset Information

The role of inverse autotransporter FdeC in Avian Pathogenic Escherichia coli adhesion

Publications

FdeC expression regulates motility and adhesion of the avian pathogenic Escherichia coli strain IMT5155.

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