Proteomic and genomic analysis reveals novel Campylobacter jejuni outer membrane proteins and potential heterogeneity

Watson, E.D.; Sherry, Aileen; Inglis, Neil F.; Lainson, Alex F.; Jyothi, Dushyanth; Yaga, Raja; Manson, Erin; Imrie, Lisa; Everest, Paul; Smith, David George Emslie

doi:10.1016/j.euprot.2014.06.003

Cited by 7 publications

(8 citation statements)

References 90 publications

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“…DMEM cultures were supplemented with 100 mM HEPES (pH 7.2). Outer membrane proteins were prepared via Sarkosyl extraction and analyzed via shotgun proteomics as previously reported ( 31 ).…”

Section: Methodsmentioning

confidence: 99%

Postgenomics Characterization of an Essential Genetic Determinant of Mammary Pathogenic Escherichia coli

Blum

Goldstone

Connolly

et al. 2018

mBio

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Escherichia coli are major bacterial pathogens causing bovine mastitis, a disease of great economic impact on dairy production worldwide. This work aimed to study the virulence determinants of mammary pathogenic E. coli (MPEC). By whole-genome sequencing analysis of 40 MPEC and 22 environmental (“dairy-farm” E. coli [DFEC]) strains, we found that only the fec locus (fecIRABCDE) for ferric dicitrate uptake was present in the core genome of MPEC and that it was absent in DFEC genomes (P < 0.05). Expression of the FecA receptor in the outer membrane was shown to be citrate dependent by mass spectrometry. FecA was overexpressed when bacteria were grown in milk. Transcription of the fecA gene and of the inner membrane transport component fecB gene was upregulated in bacteria recovered from experimental intramammary infection. The presence of the fec system was shown to affect the ability of E. coli to grow in milk. While the rate of growth in milk of fec-positive (fec+) DFEC was similar to that of MPEC, it was significantly lower in DFEC lacking fec. Furthermore, deletion of fec reduced the rate of growth in milk of MPEC strain P4, whereas fec-transformed non-mammary gland-pathogenic DFEC strain K71 gained the phenotype of the level of growth in milk observed in MPEC. The role of fec in E. coli intramammary pathogenicity was investigated in vivo in cows, with results showing that an MPEC P4 mutant lacking fec lost its ability to induce mastitis, whereas the fec+ DFEC K71 mutant was able to trigger intramammary inflammation. For the first time, a single molecular locus was shown to be crucial in MPEC pathogenicity.

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Section: Methodsmentioning

confidence: 99%

Postgenomics Characterization of an Essential Genetic Determinant of Mammary Pathogenic Escherichia coli

Blum

Goldstone

Connolly

et al. 2018

mBio

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“…; Watson et al . ). The reason for this is unknown, but as shown for other bacteria, Fno may express nonclassically associated outer membrane ‘moonlighting’ proteins on its surface, which are known to have more than one function both within the cytoplasm and extracellularly, and which have been reported to be associated with bacterial virulence (Henderson and Martin ).…”

Section: Discussionmentioning

confidence: 97%

“…LC‐ESI‐MS/MS was performed as described by Watson et al . (), with instrument parameters based on those used previously by Batycka et al . ().…”

Section: Methodsmentioning

confidence: 99%

Characterization of the outer membrane proteome ofFrancisella noatunensissubsp.orientalis

et al. 2018

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“…The C-terminal ␤-barrel domain, or autotransporter domain, is thought to form a pore in the outer membrane through which the passenger domain is secreted (20). The only autotransporters identified in C. jejuni thus far are CapA, CapB, and an additional autotransporter encoded by the CJ81176_1367 locus in C. jejuni 81-176 (22,48). CapA is an autotransporter that was previously shown to function as an auxiliary adhesin and may contribute to chicken colonization (22).…”

Section: Discussionmentioning

confidence: 99%

“…Melanization is triggered by recognition of microbial products, which are often surface exposed, by soluble pattern recognition receptors. This causes a serine protease cascade resulting in cleavage of prophenoloxidase (ProPO) to PO, which in turn, converts phenols to quinines that polymerize to form melanin (48,50). Melanin is deposited to encapsulate pathogens, resulting in pigmentation of the larvae (49,58).…”

Section: Discussionmentioning

confidence: 99%

CapC, a Novel Autotransporter and Virulence Factor of Campylobacter jejuni

Mehat

Park

Vliet

et al. 2018

Appl Environ Microbiol

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is recognized as an important causative agent of bacterial gastroenteritis in the developed world. Despite the identification of several factors contributing to infection, characterization of the virulence strategies employed by remains a significant challenge. Bacterial autotransporter proteins are a major class of secretory proteins in Gram-negative bacteria, and notably, many autotransporter proteins contribute to bacterial virulence. The aim of this study was to characterize the 81116 C8J_1278 gene (), predicted to encode an autotransporter protein, and examine the contribution of this factor to virulence of The predicted CapC protein has a number of features that are consistent with autotransporters, including the N-terminal signal sequence and the C-terminal β-barrel domain and was determined to localize to the outer membrane. Inactivation of the gene in 81116 and M1 resulted in reduced insecticidal activity in larvae. Furthermore, mutants displayed significantly reduced adherence to and invasion of nonpolarized, partially differentiated Caco-2 and T84 intestinal epithelial cells. Gentamicin treatment showed that the reduced invasion of the mutant is primarily caused by reduced adherence to intestinal epithelial cells, not by reduced invasion capability. mutants caused reduced interleukin 8 (IL-8) secretion from intestinal epithelial cells and elicited a significantly diminished immune reaction in larvae, indicating that CapC functions as an immunogen. In conclusion, CapC is a new virulence determinant of that contributes to the integral infection process of adhesion to human intestinal epithelial cells. is a major causative agent of human gastroenteritis, making this zoonotic pathogen of significant importance to human and veterinary public health worldwide. The mechanisms by which interacts with intestinal epithelial cells and causes disease are still poorly understood due, in part, to the heterogeneity of infection biology. Given the importance of to public health, the need to characterize novel and existing virulence mechanisms is apparent. The significance of our research is in demonstrating the role of CapC, a novel virulence factor in that contributes to adhesion and invasion of the intestinal epithelium, thereby in part, addressing the dearth of knowledge concerning the factors involved in pathogenesis and the variation observed in the severity of human infection.

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Proteomic and genomic analysis reveals novel Campylobacter jejuni outer membrane proteins and potential heterogeneity

Abstract: Graphical abstract

Cited by 7 publications

References 90 publications

Postgenomics Characterization of an Essential Genetic Determinant of Mammary Pathogenic Escherichia coli

Postgenomics Characterization of an Essential Genetic Determinant of Mammary Pathogenic Escherichia coli

Characterization of the outer membrane proteome ofFrancisella noatunensissubsp.orientalis

CapC, a Novel Autotransporter and Virulence Factor of Campylobacter jejuni

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