A Membrane-located Glycosyltransferase Complex Required for Biosynthesis of the d-Galactan I Lipopolysaccharide O Antigen in Klebsiella pneumoniae

Kos, Veronica; Whitfield, Chris

doi:10.1074/jbc.m110.122598

Cited by 31 publications

(26 citation statements)

References 49 publications

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“…The lipid A component of JIE4282 differed in size from all other K. pneumoniae CG258 (S1 Fig.). According to Kleborate (26), JIE4282 is missing the wbbM gene encoding a glycosyltransferase required for d -galactan I biosynthesis (43). Of note, although JIE2793 (ST512) is missing one of the hypothetical proteins ( glmA ) in the O antigen operon (O2v2 type), this had no observable impact on the observed LPS profile.…”

Section: Resultsmentioning

confidence: 99%

K. pneumoniaeST258 genomic variability and bacteriophage susceptibility

Venturini

Zakour

Bowring

et al. 2019

Preprint

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24Multidrug resistant carbapenemase-producing Klebsiella pneumoniae capable of causing 25 severe disease in humans is classified as an urgent threat by health agencies worldwide. 26 Bacteriophages are being actively explored as potential therapeutics against these multidrug 27 resistant pathogens. We are currently developing bacteriophage therapy against carbepenem-28 resistant K. pneumoniae belonging to the genetically diverse, globally disseminated clonal 29 group CG258. In an effort to define a robust experimental approach for effective selection of 30 lytic viruses for therapy, we have fully characterized the bacterial genomes of 18 target 31 strains, tested them against novel lytic bacteriophages, and generated phage-susceptibility 32 profiles. The genomes of K. pneumoniae isolates carrying blaNDM and blaKPC were sequenced 33 and isolates belonging to CG258 were selected for susceptibility testing using a panel of lytic 34 bacteriophages (n=65). The local K. pneumoniae CG258 population was dominated by 35 isolates belonging to sequence type ST258 clade 1 (86%). The primary differences between 36 ST258 genomes were variations in the capsular locus (cps) and in prophage content. We 37 showed that CG258-specific lytic phages primarily target the capsule, and that successful 38 infection is blocked in many, post-adsorption, by immunity conferred by existing prophages. 39Five bacteriophages specifically active against K. pneumoniae ST258 clade 1 (n=5) 40 belonging to the Caudovirales order were selected for further characterization. Our findings 41 show that effective control of K. pneumoniae CG258 with phage will require mixes of 42 diverse lytic viruses targeting all relevant cps variants and allowing for variable prophage 43 content. These insights will facilitate identification and selection of therapeutic phage 44 candidates against this serious pathogen. 45 46 47 48 Importance 49Bacteriophages are natural agents that exclusively and selectively kill bacteria and have the 50 potential to be useful in the treatment of multidrug resistant infections. K. pneumoniae 51 CG258 is a main agent of life-threatening sepsis that is often resistant to last-line antibiotics. 52Our work highlights some key requirements for developing bacteriophage preparations 53 targeting this pathogen. By defining the genomic profile of our clinical K. pneumoniae 54 CG258 population and matching it with bacteriophage susceptibility patterns, we found that 55 bacteriophage ability to lyse each strain correlates well with K. pneumoniae CG258 structural 56 subtypes (capsule variants). This indicates that preparation of bacteriophage therapeutics 57 targeting this pathogen should aim at including phages against each bacterial capsular 58 subtype. This necessitates a detailed understanding of the diversity of circulating isolates in 59 different geographical areas in order to make rational therapeutic choices. 60 61 62 Klebsiella pneumoniae is an important ubiquitous Gram-negative species capable of causing 63 disease in both humans an...

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Section: Resultsmentioning

confidence: 99%

K. pneumoniaeST258 genomic variability and bacteriophage susceptibility

Venturini

Zakour

Bowring

et al. 2019

Preprint

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“…The specificity of the glycosyltransferases may change when they are expressed in bacterial systems. Moreover, due to the fact that some glycosyltransferases physically interacting with each other in vivo to efficiently produce oligo-and polysaccharides [111], some glycosyltransferases expressed in bacteria may show impaired activity or specificity in the absence of their partner enzymes [107]. Therefore, additional improvements are needed to establish a cost-efficient, reliable system to produce humanized glycoproteins in E. coli or other bacteria.…”

Section: Bacterial Expression Systemsmentioning

confidence: 97%

Glycoproteins: Chemical Features and Biological Roles

Li¹,

Li²,

Chen³

2015

Carbohydrate Chemistry: State of the Art and Challenges for Drug Development

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“…Several genetic and structural studies revealed complex interrelations between proteins engaged in these processes, e.g. Wzx-Wzy/Wzz [12] , Wzy-Wzz [13] , [14] , Wzc-Wza [15] , as well as between glycosyltransferases [16] , [17] .…”

Section: Introductionmentioning

confidence: 99%

PssP2 Is a Polysaccharide Co-Polymerase Involved in Exopolysaccharide Chain-Length Determination in Rhizobium leguminosarum

et al. 2014

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Production of extracellular polysaccharides is a complex process engaging proteins localized in different subcellular compartments, yet communicating with each other or even directly interacting in multicomponent complexes. Proteins involved in polymerization and transport of exopolysaccharide (EPS) in Rhizobium leguminosarum are encoded within the chromosomal Pss-I cluster. However, genes implicated in polysaccharide synthesis are common in rhizobia, with several homologues of pss genes identified in other regions of the R. leguminosarum genome. One such region is chromosomally located Pss-II encoding proteins homologous to known components of the Wzx/Wzy-dependent polysaccharide synthesis and transport systems. The pssP2 gene encodes a protein similar to polysaccharide co-polymerases involved in determination of the length of polysaccharide chains in capsule and O-antigen biosynthesis. In this work, a mutant with a disrupted pssP2 gene was constructed and its capabilities to produce EPS and enter into a symbiotic relationship with clover were studied. The pssP2 mutant, while not altered in lipopolysaccharide (LPS), displayed changes in molecular mass distribution profile of EPS. Lack of the full-length PssP2 protein resulted in a reduction of high molecular weight EPS, yet polymerized to a longer length than in the RtTA1 wild type. The mutant strain was also more efficient in symbiotic performance. The functional interrelation between PssP2 and proteins encoded within the Pss-I region was further supported by data from bacterial two-hybrid assays providing evidence for PssP2 interactions with PssT polymerase, as well as glycosyltransferase PssC. A possible role for PssP2 in a complex involved in EPS chain-length determination is discussed.

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A Membrane-located Glycosyltransferase Complex Required for Biosynthesis of the d-Galactan I Lipopolysaccharide O Antigen in Klebsiella pneumoniae

Cited by 31 publications

References 49 publications

K. pneumoniaeST258 genomic variability and bacteriophage susceptibility

K. pneumoniaeST258 genomic variability and bacteriophage susceptibility

Glycoproteins: Chemical Features and Biological Roles

PssP2 Is a Polysaccharide Co-Polymerase Involved in Exopolysaccharide Chain-Length Determination in Rhizobium leguminosarum

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