Genetics, Biosynthesis and Assembly of O-Antigen

Valvano, Miguel A.; Furlong, Sarah E; Patel, Kinnari B.

doi:10.1007/978-3-7091-0733-1_9

Cited by 23 publications

(24 citation statements)

References 216 publications

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“…WaaL can specifically recognize one or more terminal sugars of the lipid A‐core acceptor (Feldman et al ., ), but shows no specificity for the structure of the lipid‐linked saccharide substrate (Heinrichs et al ., ). Therefore, despite different OAgs can be recombinantly expressed in Escherichia coli distinct WaaL proteins cannot cross‐complement an E. coli K‐12 waaL mutant (Valvano et al ., ). The periplasmic location of the ligation reaction suggests that potential inhibitory molecules need not enter into the bacterial cytosol and could evade membrane drug efflux mechanisms, which reduces the efficacy of conventional antibiotics.…”

Section: Introductionmentioning

confidence: 97%

Escherichia coli and Pseudomonas aeruginosa lipopolysaccharide O‐antigen ligases share similar membrane topology and biochemical properties

Ruan

Feria

Hamad

et al. 2018

Molecular Microbiology

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WaaL is an inner membrane glycosyltransferase that catalyzes the transfer of O-antigen polysaccharide from its lipid-linked intermediate to a terminal sugar of the lipid A-core oligosaccharide, a conserved step in lipopolysaccharide biosynthesis. Ligation occurs at the periplasmic side of the bacterial cell membrane, suggesting the catalytic region of WaaL faces the periplasm. Establishing the membrane topology of the WaaL protein family will enable understanding its mechanism and exploit it as a potential antimicrobial target. Applying oxidative labeling of native methionine/cysteine residues, we previously validated a topological model for Escherichia coli WaaL, which differs substantially from the reported topology of the Pseudomonas aeruginosa WaaL, derived from the analysis of truncated protein reporter fusions. Here, we examined the topology of intact E. coli and P. aeruginosa WaaL proteins by labeling engineered cysteine residues with the membrane-impermeable sulfhydryl reagent polyethylene glycol maleimide (PEG-Mal). The accessibility of PEG-Mal to targeted engineered cysteine residues in both E. coli and P. aeruginosa WaaL proteins demonstrates that both ligases share similar membrane topology. Further, we also demonstrate that P. aeruginosa WaaL shares similar functional properties with E. coli WaaL and that E. coli WaaL may adopt a functional dimer conformation.

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Section: Introductionmentioning

confidence: 97%

Escherichia coli and Pseudomonas aeruginosa lipopolysaccharide O‐antigen ligases share similar membrane topology and biochemical properties

Ruan

Feria

Hamad

et al. 2018

Molecular Microbiology

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“…1A). In the Wzx/Wzy and ABC-transporter dependent pathways, synthesis generally starts with the transfer of a sugar-1-P from a UDP-sugar to an undecaprenyl phosphate (Und-P) molecule in the inner leaflet of the inner membrane (IM) to form an Und-PP-sugar molecule (28). The priming enzymes are broadly classified in two groups, polyisoprenyl-phosphate hexose-1-phosphate transferases (PHPTs) or polyisoprenyl-phosphate N -acetylhexosamine-1-phosphate transferases (PNPTs) (29).…”

Section: Introductionmentioning

confidence: 99%

Characterization of the exopolysaccharide biosynthesis pathway inMyxococcus xanthus

Pérez‐Burgos

García‐Romero

Jung

et al. 2020

Preprint

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18Myxococcus xanthus arranges into two morphologically distinct biofilms depending on its 19 nutritional status, i.e. coordinately spreading colonies in the presence of nutrients and spore-20 filled fruiting bodies in the absence of nutrients. A secreted polysaccharide referred to as 21 exopolysaccharide (EPS) is a structural component of both biofilms and is also important for 22Here, we characterized the EPS biosynthetic machinery and show that it makes up a 45 Wzx/Wzy-dependent pathway for polysaccharide biosynthesis. Mutants lacking a component 46 of this pathway had reduced type IV pili-dependent motility and a conditional defect in 47 development. Also, these analysis suggest that EPS and/or the EPS biosynthetic machinery 48 is important for type IV pili formation. 49 50 51

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“…There are four distinct processes for synthesis and translocation of the O antigen, i.e. the Wzx/Wzy-, ATP-binding cassette (ABC) transporter-, synthase-and Wzk-dependent processes (Valvano et al, 2011). Among them, the first pathway occurs in the synthesis of the majority of O antigens, especially in heteropolymeric O antigens.…”

Section: Serological Relationships Are Observed Between Differentmentioning

confidence: 99%

“…O unit synthesis is initiated by transfer of the first sugar 1-phosphate from the UDP precursor to the lipid acceptor undecaprenyl phosphate (UndP), catalysed by WecA (Valvano et al, 2011). In Enterobacteriaceae, the wecA gene is located in the ECA gene cluster rather than the OGC (Valvano, 2003).…”

Section: Sugar Transferase Genesmentioning

confidence: 99%

“…The wzx and wzy genes were found in all eight Providencia OGCs studied. The topology models of the putative Wzx proteins derived from in silico prediction contain 10-14 well-proportioned transmembrane segments, which is typical of O unit flippases (Valvano et al, 2011). The putative Wzy proteins have 9-12 predicted transmembrane segments, with a large periplasmic loop of 36-88 amino acid residues.…”

Section: O Antigen Processing Genesmentioning

confidence: 99%

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Localization and molecular characterization of putative O antigen gene clusters of Providencia species

et al. 2012

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Enterobacteria of the genus Providencia are opportunistic human pathogens associated with urinary tract and wound infections, as well as enteric diseases. The lipopolysaccharide (LPS) O antigen confers major antigenic variability upon the cell surface and is used for serotyping of Gram-negative bacteria. Recently, Providencia O antigen structures have been extensively studied, but no data on the location and organization of the O antigen gene cluster have been reported. In this study, the four Providencia genome sequences available were analysed, and the putative O antigen gene cluster was identified in the polymorphic locus between the cpxA and yibK genes. This finding provided the necessary information for designing primers, and cloning and sequencing the O antigen gene clusters from five more Providencia alcalifaciens strains. The gene functions predicted in silico were in agreement with the known O antigen structures; furthermore, annotation of the genes involved in the three-step synthesis of GDP-colitose (gmd, colD and colC) was supported by cloning and biochemical characterization of the corresponding enzymes. In one strain (P. alcalifaciens O39), no polysaccharide product of the gene cluster in the cpxA-yibK locus was found, and hence genes for synthesis of the existing O antigen are located elsewhere in the genome. In addition to the putative O antigen synthesis genes, homologues of wza, wzb, wzc and (in three strains) wzi, required for the surface expression of capsular polysaccharides, were found upstream of yibK in all species except Providencia rustigianii, suggesting that the LPS of these species may be attributed to the so-called K LPS (K LPS ). The data obtained open a way for development of a PCR-based typing method for identification of Providencia isolates. INTRODUCTIONThe genus Providencia of the family Enterobacteriaceae currently consists of eight species (Juneja & Lazzaro, 2009;Somvanshi et al., 2006;O'Hara et al., 2000), among which Providencia stuartii, Providencia rettgeri, Providencia rustigianii and Providencia alcalifaciens are the species that most commonly cause human infection. P. stuartii is often found in complicated urinary tract infections in patients with chronic indwelling urinary catheters (Rahav et al., 1994). Providencia species, especially P. rustigianii and P. alcalifaciens, are an infrequent cause of traveller's diarrhea (Yoh 3These authors contributed equally to this work.Abbreviations: 6dTal, 6-deoxytalose; Col, colitose; CPS, capsular polysaccharide; ECA, enterobacterial common antigen; ESI-MS, electrospray ionization-MS; Fuc3NAc, 3-acetamido-3,6-dideoxygalactose; Kdo, 3-deoxy-D-manno-oct-2-ulosonic acid; OGC, O antigen gene cluster; PLP, pyridoxal 59-phosphate; Qui, quinovose; TOCSY, total correlation spectroscopy; UndPP, undecaprenyl diphosphate.The GenBank/EMBL/DDBJ accession numbers for the cpxA-yibK region sequences of P. alcalifaciens O28, O36, O44 and O39 are JQ319041, HM583639, JN097784 and JN097785, respectively.Three supplementary figures and three supplem...

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Genetics, Biosynthesis and Assembly of O-Antigen

Cited by 23 publications

References 216 publications

Escherichia coli and Pseudomonas aeruginosa lipopolysaccharide O‐antigen ligases share similar membrane topology and biochemical properties

Escherichia coli and Pseudomonas aeruginosa lipopolysaccharide O‐antigen ligases share similar membrane topology and biochemical properties

Characterization of the exopolysaccharide biosynthesis pathway inMyxococcus xanthus

Localization and molecular characterization of putative O antigen gene clusters of Providencia species

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