The
            <i>Klebsiella pneumoniae wabG</i>
            Gene: Role inBiosynthesis of the Core Lipopolysaccharide andVirulence

Izquierdo, Luís; Coderch, Núria; Piqué, Núria; Bedini, Emiliano; Corsaro, Maria Michela; Merino, Susana; Fresno, Sandra; Tomás, Juan M.; Regué, Miguel

doi:10.1128/jb.185.24.7213-7221.2003

Cited by 83 publications

(94 citation statements)

References 33 publications

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“…Thus, it is predicted that these strains contain wabG, wabH, and wabN homologues. This has been shown for S. marcescens N28b where these gene homologues are able to complement the corresponding K. pneumoniae mutants as shown in this and previous work (7,11,17). Interestingly these three genes are also found as predicted in P. mirabilis HI4320 (pedant.gsf.de/cgi-bin/wwwfly.pl?SetϭProteus_mirabilis& Pageϭindex).…”

Section: Discussionsupporting

confidence: 50%

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The Incorporation of Glucosamine into Enterobacterial Core Lipopolysaccharide

Regué

Izquierdo

Fresno

et al. 2005

Journal of Biological Chemistry

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The core lipopolysaccharide (LPS) of Klebsiella pneumoniae is characterized by the presence of disaccharide ␣GlcN-(1,4)-␣GalA attached by an ␣1,3 linkage to L-glycero-D-manno-heptopyranose II (LD-HeppII). Previously it has been shown that the WabH enzyme catalyzes the incorporation of GlcNAc from UDP-GlcNAc to outer core LPS. The presence of GlcNAc instead of GlcN and the lack of UDP-GlcN in bacteria indicate that an additional enzymatic step is required. In this work we identified a new gene (wabN) in the K. pneumoniae core LPS biosynthetic cluster. Chemical and structural analysis of K. pneumoniae non-polar wabN mutants showed truncated core LPS with GlcNAc instead of GlcN. In vitro assays using LPS truncated at the level of D-galacturonic acid (GalA) and cell-free extract containing WabH and WabN together led to the incorporation of GlcN, whereas none of them alone were able to do it. This result suggests that the later enzyme (WabN) catalyzes the deacetylation of the core LPS containing the GlcNAc residue. Thus, the incorporation of the GlcN residue to core LPS in K. pneumoniae requires two distinct enzymatic steps. WabN homologues are found in Serratia marcescens and some Proteus strains that show the same disaccharide ␣GlcN-(1,4)-␣GalA attached by an ␣1,3 linkage to LD-HeppII.

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Section: Discussionsupporting

confidence: 50%

“…1). Furthermore the functions in both inner and outer core biosynthesis of most of the transferases encoded by the wa gene products have been elucidated (7)(8)(9)(10)(11)(12) (Fig. 1).…”

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confidence: 99%

The Incorporation of Glucosamine into Enterobacterial Core Lipopolysaccharide

Regué

Izquierdo

Fresno

et al. 2005

Journal of Biological Chemistry

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“…The objective of this study was to examine the biosynthesis of this novel region of the core OS. The gene product involved in the addition of the GalUA residue (WabG) was previously identified by others (10), and the remaining transferases were established in this present study.…”

Section: Discussionmentioning

confidence: 70%

“…Genes responsible for inner core synthesis are readily identified by conserved sequences shared with E. coli and Salmonella, and the enzymology of the biosynthesis of the K. pneumoniae Hep-Kdo inner core backbone has been verified (13). Preliminary structural analysis of a non-polar waaE mutant indicates that WaaE is involved in the addition of the inner core ␤-Glc residue (14) and that WabG is required for the addition of the initial GalUA residue of the outer core (10). The functions of the remaining transferases have not been assigned.…”

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Biosynthesis of a Novel 3-Deoxy-D-manno-oct-2-ulosonic Acid-containing Outer Core Oligosaccharide in the Lipopolysaccharide of Klebsiella pneumoniae

Frirdich

Vinogradov

Whitfield

2004

Journal of Biological Chemistry

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The core oligosaccharide region of Klebsiella pneumoniae lipopolysaccharide contains some novel features that distinguish it from the corresponding lipopolysaccharide region in other members of the Enterobacteriaceae family, such as Escherichia coli and Salmonella. The conserved Klebsiella outer core contains the unusual trisaccharide 3-deoxy-D-manno-oct-2-ulosonic acid (Kdo)-(2,6)-GlcN-(1,4)-GalUA. In general, Kdo residues are normally found in the inner core, but in K. pneumoniae, this Kdo residue provides the ligation site for O polysaccharide. The outer core Kdo residue can also be non-stoichiometrically substituted with an L-glycero-D-manno-heptopyranose (Hep) residue, another component more frequently found in the inner core. To understand the genetics and biosynthesis of core oligosaccharide synthesis in Klebsiella, the gene products involved in the addition of the outer core GlcN (WabH), Kdo (WabI), and Hep (WabJ) residues as well as the inner core HepIII residue (WaaQ) were identified. Non-polar mutations were created in each of the genes, and the resulting mutant lipopolysaccharide was analyzed by mass spectrometry. The in vitro glycosyltransferase activity of WabI and WabH was verified. WabI transferred a Kdo residue from CMP-Kdo onto the acceptor lipopolysaccharide. The activated precursor required for GlcN addition has not been identified. However, lysates overexpressing WabH were able to transfer a GlcNAc residue from UDP-GlcNAc onto the acceptor GalUA residue in the outer core.

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Characterization of the Core Oligosaccharide and the O‐Antigen Biological Repeating Unit from Halomonas stevensii Lipopolysaccharide: The First Case of O‐Antigen Linked to the Inner Core

Pieretti

Carillo

Lindner

et al. 2012

Chemistry A European J

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A novel core structure among bacterial lipopolysaccharides (LPS) that belong to the genus Halomonas has been characterized. H. stevensii is a moderately halophilic microorganism, as are the majority of the Halomonadaceae. It brought to light the pathogenic potential of this genus. On account of their role in immune system elicitation, elucidation of LPS structure is the mandatory starting point for a deeper understanding of the interaction mechanisms between host and pathogen. In this paper we report the structure of the complete saccharidic portion of the LPS from H. stevensii. In contrast to the finding that the O-antigen is usually covalently linked to the outer core oligosaccharide, we could demonstrate that the O-polysaccharide of H. stevensii is linked to the inner core of an LPS. By means of high-performance anion-exchange chromatography with pulsed amperometric detection we were able to isolate the core decasaccharide as well as a tridecasaccharide constituted by the core region plus one O-repeating unit after alkaline degradation of the LPS. The structure was elucidated by one- and two-dimensional NMR spectroscopy, ESI Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry, and chemical analysis.

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The Klebsiella pneumoniae wabG Gene: Role inBiosynthesis of the Core Lipopolysaccharide andVirulence

Cited by 83 publications

References 33 publications

The Incorporation of Glucosamine into Enterobacterial Core Lipopolysaccharide

The Incorporation of Glucosamine into Enterobacterial Core Lipopolysaccharide

Biosynthesis of a Novel 3-Deoxy-D-manno-oct-2-ulosonic Acid-containing Outer Core Oligosaccharide in the Lipopolysaccharide of Klebsiella pneumoniae

Characterization of the Core Oligosaccharide and the O‐Antigen Biological Repeating Unit from Halomonas stevensii Lipopolysaccharide: The First Case of O‐Antigen Linked to the Inner Core

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