Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism

Mann, Evan; Ovchinnikova, Olga G.; King, Jerry D.; Whitfield, Chris

doi:10.1074/jbc.m115.660803

Cited by 22 publications

(20 citation statements)

References 55 publications

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“…Terminating ␤-Kdo Residue Is Required for O-PS ExportBiosynthesis of the O12 OPS requires glycosyltransferase (GT) activity provided by the WbbL and WbbB proteins (26). WbbB contains three predicted GT catalytic domains.…”

Section: Resultsmentioning

confidence: 99%

“…2A). The Western immunoblot detects both LPS and unexported Und-PP-linked glycan, whereas the silver-stained gel only reports O-PS that is exported and incorporated into LPS (26). The uncapped O12 glycan was still exported by the O2a ABC transporter, as evidenced in the silver-stained SDS-PAGE, but it was no longer a substrate for the native O12 ABC transporter.…”

Section: Resultsmentioning

confidence: 99%

“…LPS was visualized with silver staining (33). Immunoblot detection of O-antigens was performed by transferring lysates separated by SDS-PAGE to nitrocellulose membranes and probing with rabbit antiserum specific for the R. terrigena ATCC 33257 O-antigen structure (26). Alkaline phosphatase-conjugated goat anti-rabbit secondary antibodies (Cedar Lane) were used, and the immunoblot was developed with 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium (Roche Applied Science).…”

Section: Methodsmentioning

confidence: 99%

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The Klebsiella pneumoniae O12 ATP-binding Cassette (ABC) Transporter Recognizes the Terminal Residue of Its O-antigen Polysaccharide Substrate

Mann

Mallette

Clarke

et al. 2016

Journal of Biological Chemistry

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Export of the Escherichia coli serotype O9a O-antigenic polysaccharides (O-PS) involves an ATP-binding cassette (ABC)transporter. The process requires a non-reducing terminal residue, which is recognized by a carbohydrate-binding module (CBM) appended to the C terminus of the nucleotide-binding domain of the transporter. Here, we investigate the process in Klebsiella pneumoniae serotype O12 (and Raoultella terrigena ATCC 33257). The O12 polysaccharide is terminated at the non-reducing end by a ␤-linked 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo) residue. The O12 ABC transporter also binds its cognate O-PS via a CBM, and export is dependent on the presence of the terminal ␤-Kdo residue. The overall structural architecture of the O12 CBM resembles the O9a prototype, but they share only weak sequence similarity, and the putative binding pocket for the O12 glycan is different. Removal of the CBM abrogated O-PS transport, but export was restored when the CBM was expressed in trans with the mutant CBM-deficient ABC transporter. These results demonstrate that the CBM-mediated substrate-recognition mechanism is evolutionarily conserved and can operate with glycans of widely differing structures.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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The Klebsiella pneumoniae O12 ATP-binding Cassette (ABC) Transporter Recognizes the Terminal Residue of Its O-antigen Polysaccharide Substrate

Mann

Mallette

Clarke

et al. 2016

Journal of Biological Chemistry

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“…Thus, the evidence suggests that the O8 LPS type has emerged by modification of the O-antigen component of the O1 LPS. O-antigen modification is a well-recognized phenomenon in Gram-negative bacteria and may be generated by horizontal gene transfer (43,44) or bacteriophage-mediated mechanisms (45,46). Indeed, horizontal gene transfer involving O-antigen biosynthesis gene clusters has contributed to LPS diversity in the Aeromonas hydrophila strains responsible for septicemia in catfish (47).…”

Section: Discussionmentioning

confidence: 99%

Yersinia ruckeri Isolates Recovered from Diseased Atlantic Salmon (Salmo salar) in Scotland Are More Diverse than Those from Rainbow Trout (Oncorhynchus mykiss) and Represent Distinct Subpopulations

Ormsby

Caws

Burchmore

et al. 2016

Appl Environ Microbiol

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Yersinia ruckeri is the etiological agent of enteric redmouth (ERM) disease of farmed salmonids. Enteric redmouth disease is traditionally associated with rainbow trout (Oncorhynchus mykiss, Walbaum), but its incidence in Atlantic salmon (Salmo salar) is increasing. Yersinia ruckeri isolates recovered from diseased Atlantic salmon have been poorly characterized, and very little is known about the relationship of the isolates associated with these two species. Phenotypic approaches were used to characterize 109 Y. ruckeri isolates recovered over a 14-year period from infected Atlantic salmon in Scotland; 26 isolates from infected rainbow trout were also characterized. Biotyping, serotyping, and comparison of outer membrane protein profiles identified 19 Y. ruckeri clones associated with Atlantic salmon but only five associated with rainbow trout; none of the Atlantic salmon clones occurred in rainbow trout and vice versa. These findings suggest that distinct subpopulations of Y. ruckeri are associated with each species. A new O serotype (designated O8) was identified in 56 biotype 1 Atlantic salmon isolates and was the most common serotype identified from 2006 to 2011 and in 2014, suggesting an increased prevalence during the time period sampled. Rainbow trout isolates were represented almost exclusively by the same biotype 2, serotype O1 clone that has been responsible for the majority of ERM outbreaks in this species within the United Kingdom since the 1980s. However, the identification of two biotype 2, serotype O8 isolates in rainbow trout suggests that vaccines containing serotypes O1 and O8 should be evaluated in both rainbow trout and Atlantic salmon for application in Scotland. IMPORTANCEVaccination plays an important role in protecting Atlantic salmon against the bacterial pathogen Yersinia ruckeri, but, in recent years, there has been an increasing incidence of vaccine breakdown in salmon. This is largely because current vaccines are aimed at rainbow trout and are based on serotypes specific for this species. A wider range of serotypes is responsible for infection in Atlantic salmon, but very little is known about the diversity of these strains and their relationships to those recovered from rainbow trout. In the present study, we demonstrate that Y. ruckeri isolates recovered from diseased Atlantic salmon in Scotland are more diverse than those from rainbow trout; furthermore, isolates from the two species represent distinct subpopulations. In addition, a new O serotype was identified that is responsible for a significant proportion of the disease in Atlantic salmon. Our findings are likely to have important implications for the development of improved vaccines against Y. ruckeri. The Gram-negative bacterium Yersinia ruckeri is the etiological agent of enteric redmouth (ERM) disease of cultured salmonids and causes significant economic losses to the fish-farming industry. Yersinia ruckeri was first isolated in 1956 from diseased rainbow trout (Oncorhynchus mykiss, Walbaum) in the Hagerman Valle...

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“…Recently, it has been demonstrated that there are no mechanistic barriers for bacteriophage-mediated O-antigen glycosylation in the ABC-transporter-dependent pathway (Mann et al, 2015). Bioinformatics data suggest that decoration of the galactan O antigens of K. pneumoniae O2 and O9 and related Raoultella terrigena by side-chain galactose residues could be natural examples of bacteriophage-mediated galactosylation .…”

Section: Discussionmentioning

confidence: 99%

O antigen of Franconibacter pulverisG3872 (O1) is a 4-deoxy-d-arabino-hexose-containing polysaccharide synthesized by the ABC-transporter-dependent pathway

Wang

Arbatsky

et al. 2016

Microbiology

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Franconibacter (Enterobacter, Cronobacter) pulveris bacteria share several typical characteristics with, and hence pose a challenge for the detection of, Cronobacter sakazakii, an emerging opportunistic pathogen, which can cause severe infections in neonates. A structurally variable O-specific polysaccharide (OPS) called O antigen provides the major basis for the typing of Gram-negative bacteria. We investigated the structure and genetics of the O antigen of F. pulveris G3872 (designated O1). An OPS was isolated by mild alkaline degradation of the LPS, whereas the same polysaccharide and its oligosaccharide fragments were obtained by mild acid degradation. Studies by sugar analysis and NMR spectroscopy showed that the OPS contained D-ribose, L-rhamnose (L-Rha) and a rarely occurring monosaccharide 4-deoxy-Darabino-hexose, and the OPS structure was established. The O-antigen gene cluster of F. pulveris G3872 between JUMPStart and gnd genes includes putative genes for glycosyltransferases, ATP-binding cassette (ABC)-transporter genes wzm and wzt, and genes for the synthesis of L-Rha, but no genes for the synthesis of 4-deoxy-D-arabino-hexose. A mutation test with the wzm gene confirmed that the OPS is synthesized and exported by the ABC-transporter-dependent pathway. A trifunctional transferase was suggested to catalyse formation of two glycosidic linkages and add a methyl group to the non-reducing end of the OPS to terminate the chain elongation. A carbohydrate-binding module that presumably recognizes the terminal methyl-modified monosaccharide was found at the C-terminus of Wzt. Primers specific for F. pulveris G3872 were designed based on the wzm gene, which has potential to be used for identification and detection of the O1 serogroup.

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Bacteriophage-mediated Glucosylation Can Modify Lipopolysaccharide O-Antigens Synthesized by an ATP-binding Cassette (ABC) Transporter-dependent Assembly Mechanism

Cited by 22 publications

References 55 publications

The Klebsiella pneumoniae O12 ATP-binding Cassette (ABC) Transporter Recognizes the Terminal Residue of Its O-antigen Polysaccharide Substrate

The Klebsiella pneumoniae O12 ATP-binding Cassette (ABC) Transporter Recognizes the Terminal Residue of Its O-antigen Polysaccharide Substrate

Yersinia ruckeri Isolates Recovered from Diseased Atlantic Salmon (Salmo salar) in Scotland Are More Diverse than Those from Rainbow Trout (Oncorhynchus mykiss) and Represent Distinct Subpopulations

O antigen of Franconibacter pulverisG3872 (O1) is a 4-deoxy-d-arabino-hexose-containing polysaccharide synthesized by the ABC-transporter-dependent pathway

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