An aflagellate mutant Yersinia enterocolitica biotype 1A strain displays altered invasion of epithelial cells, persistence in macrophages, and cytokine secretion profiles in vitro

McNally, Alan; Ragione, Roberto M. La; Best, Angus; Manning, Georgina; Newell, Diane G.

doi:10.1099/mic.0.2006/000919-0

Cited by 27 publications

(30 citation statements)

References 46 publications

(36 reference statements)

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“…All mutagenesis studies were performed in the genome-sequenced type PG 1 strain YE5303 (McNally et al, 2007;Reuter et al, 2014). A cytolethal distending toxin (CDT) mutant was made by PCR, amplifying cdtB using primers CDTFor and CDTRev (Table 2), and cloning into pCRTopo2.1 (Invitrogen) to create pAD5.…”

Section: Methodsmentioning

confidence: 99%

“…The inactivated cdtB gene was then PCR amplified and subcloned into the SmaI site of pKNG101. The resulting plasmid was used to transform E. coli S17-1 Pir cells, and these were used as donor cells in a filter mating conjugation with YE5303 (McNally et al, 2007). A functional Yersinia genus type III secretion system (YGT) mutant was constructed by PCR, amplifying the apparatus encoding gene ygtV (Reuter et al, 2014) using primers ygtvFor and ygtvRev (Table 2) and cloning into pCRTopo2.1 to create pAR1.…”

Section: Methodsmentioning

confidence: 99%

“…The inactivated ygtV gene was then PCR amplified and subcloned into the SmaI site of pKNG101. The resulting plasmid, pAR3, was used to transform E. coli S17-1 Pir cells, and these were used as donor cells in a filter mating conjugation with YE5303 (McNally et al, 2007). Complementation of the cdtB and ygtV mutants was performed by transforming the mutated strains with pAD5 and pAR1, respectively.…”

Section: Methodsmentioning

confidence: 99%

“…It is known that many PG 1 isolates carry genes proposed to be Yersinia virulence factors (Kumar & Virdi, 2012;Singh & Virdi, 2004;Tennant et al, 2005) and population genomic studies have shown that many genes purported to play a role in Y. enterocolitica pathogenesis of other PGs are found in PG 1 isolates; additionally a putative type III secretion system is found exclusively in PG 1 (Reuter et al, 2014). To date, the only factor that has been shown to be involved in a virulence-associated trait in PG 1 is the requirement of flagella to survive inside macrophages (McNally et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

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All Yersinia enterocolitica are pathogenic: virulence of phylogroup 1 Y. enterocolitica in a Galleria mellonella infection model

et al. 2016

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Yersinia enterocolitica is a zoonotic pathogen and a common cause of gastroenteritis in humans. The species is composed of six diverse phylogroups, of which strains of phylogroup 1 are considered non-pathogenic to mammals due to the lack of the major virulence plasmid pYV, and their lack of virulence in a mouse infection model. In the present report we present data examining the pathogenicity of strains of Y. enterocolitica across all six phylogroups in a Galleria mellonellla model. We have demonstrated that in this model strains of phylogroup 1 exhibit severe pathogenesis with a lethal dose of as low as 10 c.f.u., that this virulence is an active process and that flagella play a major role in the virulence phenotype. We have also demonstrated that the complete lack of virulence in Galleria of the mammalian pathogenic phylogroups is not due to carriage of the pYV virulence plasmid. Our data suggest that all Y. enterocolitica can be pathogenic, which may be a reflection of the true natural habitat of the species, and that we may need to reconsider the eco-evo perspective of this important bacterial species.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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All Yersinia enterocolitica are pathogenic: virulence of phylogroup 1 Y. enterocolitica in a Galleria mellonella infection model

et al. 2016

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“…LOS was extracted using a modification of the phenol-water extraction procedure (32,40). Briefly, confluent bacterial growth from agar plates was washed three times in PBS (Sigma, United Kingdom), and cells were resuspended in sterile distilled water to an optical density at 600 nm (OD 600 ) of 1.0.…”

Section: Methodsmentioning

confidence: 99%

Cj1136 Is Required for Lipooligosaccharide Biosynthesis, Hyperinvasion, and Chick Colonization by Campylobacter jejuni

et al. 2012

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ABSTRACTCampylobacter jejuniis a major cause of bacterial food-borne enteritis worldwide, and invasion into intestinal epithelial cells is an important virulence mechanism. Recently we reported the identification of hyperinvasiveC. jejunistrains and created a number of transposon mutants of one of these strains, some of which exhibited reduced invasion into INT-407 and Caco-2 cells. In one such mutant the transposon had inserted into a homologue ofcj1136, which encodes a putative galactosyltransferase according to the annotation of theC. jejuniNCTC11168 genome. In the current study, we investigated the role ofcj1136inC. jejunivirulence, lipooligosaccharide (LOS) biosynthesis, and host colonization by targeted mutagenesis and complementation of the mutation. Thecj1136mutant showed a significant reduction in invasion into human intestinal epithelial cells compared to the wild-type strain 01/51. Invasion levels were partially restored on complementing the mutation. The inactivation ofcj1136resulted in the production of truncated LOS, while biosynthesis of a full-length LOS molecule was restored in the complemented strain. Thecj1136mutant showed an increase in sensitivity to the bile salts sodium taurocholate and sodium deoxycholate and significantly increased sensitivity to polymyxin B compared to the parental strain. Importantly, the ability of the mutant to colonize 1-day-old chicks was also significantly impaired. This study confirms that a putative galactosyltransferase encoded bycj1136is involved in LOS biosynthesis and is important forC. jejunivirulence, as disruption of this gene and the resultant truncation of LOS affect both colonizationin vivoand invasivenessin vitro.

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Yersinia spp.

Johnson¹

2017

Bacterial Pathogens and Their Virulence Factors

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An aflagellate mutant Yersinia enterocolitica biotype 1A strain displays altered invasion of epithelial cells, persistence in macrophages, and cytokine secretion profiles in vitro

Cited by 27 publications

References 46 publications

All Yersinia enterocolitica are pathogenic: virulence of phylogroup 1 Y. enterocolitica in a Galleria mellonella infection model

All Yersinia enterocolitica are pathogenic: virulence of phylogroup 1 Y. enterocolitica in a Galleria mellonella infection model

Cj1136 Is Required for Lipooligosaccharide Biosynthesis, Hyperinvasion, and Chick Colonization by Campylobacter jejuni

Yersinia spp.

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