The failure of different strains of<i>Yersinia pestis</i>to produce lipopolysaccharide O-antigen under different growth conditions is due to mutations in the O-antigen gene cluster

Prior, Joann L.; Parkhill, Julian; Hitchen, Paul G.; Mungall, Karen; Stevens, Kim; Morris, Howard R.; Reason, Andrew J.; Oyston, Petra C. F.; Dell, Anne; Wren, Brendan W.; Titball, Richard W.

doi:10.1111/j.1574-6968.2001.tb10608.x

Cited by 55 publications

(18 citation statements)

References 24 publications

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“…In Y. pestis , the LOS is composed of a short carbohydrate (oligosaccharide core) chain bound to lipid A. O antigen is not present in Y. pestis due to frameshift mutations in the ddhB , gmd , fcl , and ushA genes (28). The lipid A portion of LPS anchors the molecule in the outer membrane of Gram-negative bacteria and is also the portion of the molecule recognized by the TLR4 complex signaling component MD2.…”

Section: Resultsmentioning

confidence: 99%

Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery

Gregg

Harberts

Gardner

et al. 2017

mBio

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Adjuvant properties of bacterial cell wall components like MPLA (monophosphoryl lipid A) are well described and have gained FDA approval for use in vaccines such as Cervarix. MPLA is the product of chemically modified lipooligosaccharide (LOS), altered to diminish toxic proinflammatory effects while retaining adequate immunogenicity. Despite the virtually unlimited number of potential sources among bacterial strains, the number of useable compounds within this promising class of adjuvants are few. We have developed bacterial enzymatic combinatorial chemistry (BECC) as a method to generate rationally designed, functionally diverse lipid A. BECC removes endogenous or introduces exogenous lipid A-modifying enzymes to bacteria, effectively reprogramming the lipid A biosynthetic pathway. In this study, BECC is applied within an avirulent strain of Yersinia pestis to develop structurally distinct LOS molecules that elicit differential Toll-like receptor 4 (TLR4) activation. Using reporter cell lines that measure NF-κB activation, BECC-derived molecules were screened for the ability to induce a lower proinflammatory response than Escherichia coli LOS. Their structures exhibit varied, dose-dependent, TLR4-driven NF-κB activation with both human and mouse TLR4 complexes. Additional cytokine secretion screening identified molecules that induce levels of tumor necrosis factor alpha (TNF-α) and interleukin-8 (IL-8) comparable to the levels induced by phosphorylated hexa-acyl disaccharide (PHAD). The lead candidates demonstrated potent immunostimulation in mouse splenocytes, human primary blood mononuclear cells (PBMCs), and human monocyte-derived dendritic cells (DCs). This newly described system allows directed programming of lipid A synthesis and has the potential to generate a diverse array of TLR4 agonist candidates.

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Section: Resultsmentioning

confidence: 99%

Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery

Gregg

Harberts

Gardner

et al. 2017

mBio

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“…Differences in the composition of the lipopolysaccharide (LPS) coating of the two species may also play a role in differing virulence. Due to multiple mutations in the O-antigen gene cluster, Y. pestis fails to produce O-antigen, a structural component of lipopolysaccharide of Gram-negative bacteria (Prior et al, 2001 ). An important step of LPS biosynthesis is the 3-deoxy-D-manno-octulosonic acid (Kdo) glycosylation of lipid A, which is catalyzed by the Kdo transferase WaaA (Tan and Darby, 2005 ).…”

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confidence: 99%

Transcriptional regulation of the waaAE-coaD operon by PhoP and RcsAB in Yersinia pestis biovar Microtus

et al. 2014

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“…The composition of the acyl chains attached to the core oligosaccharide also varies with temperature [17]. Y. pestis CO92 LPS does not contain O-antigen at either temperature [43]. …”

Section: Methodsmentioning

confidence: 99%

An experimentally-supported genome-scale metabolic network reconstruction for Yersinia pestis CO92

et al. 2011

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BackgroundYersinia pestis is a gram-negative bacterium that causes plague, a disease linked historically to the Black Death in Europe during the Middle Ages and to several outbreaks during the modern era. Metabolism in Y. pestis displays remarkable flexibility and robustness, allowing the bacterium to proliferate in both warm-blooded mammalian hosts and cold-blooded insect vectors such as fleas.ResultsHere we report a genome-scale reconstruction and mathematical model of metabolism for Y. pestis CO92 and supporting experimental growth and metabolite measurements. The model contains 815 genes, 678 proteins, 963 unique metabolites and 1678 reactions, accurately simulates growth on a range of carbon sources both qualitatively and quantitatively, and identifies gaps in several key biosynthetic pathways and suggests how those gaps might be filled. Furthermore, our model presents hypotheses to explain certain known nutritional requirements characteristic of this strain.ConclusionsY. pestis continues to be a dangerous threat to human health during modern times. The Y. pestis genome-scale metabolic reconstruction presented here, which has been benchmarked against experimental data and correctly reproduces known phenotypes, provides an in silico platform with which to investigate the metabolism of this important human pathogen.

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The failure of different strains ofYersinia pestisto produce lipopolysaccharide O-antigen under different growth conditions is due to mutations in the O-antigen gene cluster

Cited by 55 publications

References 24 publications

Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery

Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery

Transcriptional regulation of the waaAE-coaD operon by PhoP and RcsAB in Yersinia pestis biovar Microtus

An experimentally-supported genome-scale metabolic network reconstruction for Yersinia pestis CO92

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