KpsF Is the Arabinose-5-phosphate Isomerase Required for 3-Deoxy-d-manno-octulosonic Acid Biosynthesis and for Both Lipooligosaccharide Assembly and Capsular Polysaccharide Expression in Neisseria meningitidis

Tzeng, Yih-Ling; Datta, Anup; Strole, Christy; Kolli, V. S. Kumar; Birck, Matthew R.; Taylor, W. P.; Carlson, Russell W.; Woodard, Ronald W.; Stephens, David S.

doi:10.1074/jbc.m200931200

Cited by 56 publications

(41 citation statements)

References 58 publications

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“…The gene for KDO 8-P phosphatase (yrbI) was recently located on the yrb gene cluster (17), and its position next to an ORF (yrbH) encoding a highly conserved putative phosphosugar isomerase suggested the possible role of API for yrbH. In addition, yrbH shares significant homology with NMB0352 in N. meningitidis serogroup B (MC58) that had been identified as encoding a protein with API activity (Table I) (25). This is the only protein in N. meningitidis that resembles either of the two paralogues in E. coli K-12.…”

Section: Discussionmentioning

confidence: 99%

“…2). NMB0352 was recently identified as encoding a protein with API activity (25). Because yrbH is located next to an enzyme involved in KDO synthesis, is highly conserved in Gram-negative bacteria, is predicted to be a phosphosugar isomerase, and has the highest degree of homology among the E. coli paralogues with NMB0352, the yrbH ORF was chosen as the candidate E. coli K-12 gene encoding for the API involved in the generation of A5P for LPS synthesis.…”

Section: Identification Of Yrbh As Api In E Coli K-12-tomentioning

confidence: 99%

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Escherichia coli YrbH Is a D-Arabinose 5-Phosphate Isomerase

Meredith

Woodard

2003

Journal of Biological Chemistry

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The lipopolysaccharide (LPS) 1 layer is an essential outer membrane glycolipid located on the cellular surface of virtually all Gram-negative bacteria (1). The LPS contains three principle elements: lipid A, the core oligosaccharide, and the Oantigen. The core oligosaccharide provides the link between the highly conserved membrane-imbedded lipid A and the structurally diverse complex carbohydrate O-antigen chain (2) and can be further subdivided into the inner and outer core regions. Whereas the structure of the outer core is somewhat variable, the inner core is highly conserved, being primarily composed of L-glycero-D-manno-heptose and 3-deoxy-D-manno-octulosonate (KDO) residues (3). This suggests the importance of the inner core-lipid A complex in maintaining the structural integrity and viability of the cell (4). In addition, it has been shown that the minimal LPS required to sustain growth in mutant Escherichia coli strains consists of the KDO 2 -lipid A core (Re endotoxin) (5, 6), that the interruption of KDO biosynthesis leads to the arrest of cell growth (7-9), and that cells containing a compromised LPS are generally less pathogenic and more susceptible to antibiotics (10, 11). Accordingly, enzymes involved in the biosynthesis of KDO are attractive targets for the development of specific antibiotics because of their localization to and high level of conservation within Gram-negative bacteria.The synthesis and activation of KDO involves four sequentially acting enzymes: D-arabinose 5-phosphate isomerase (API), deoxy-D-manno-octulosonate 8-phosphate (KDO 8-P) synthase, KDO 8-P phosphatase, and cytidine 5Ј-monophosphate-KDO synthetase ( Fig. 1) (12, 13). The first committed step to KDO generation is catalyzed by 3-deoxy-D-manno-octulosonate 8-phosphate (KDO 8-P) synthase, which condenses D-arabinose 5-phosphate (A5P) with phosphoenolpyruvate via a stereospecific aldol-type condensation (14, 15). A5P is the first intermediate unique to the KDO biosynthetic pathway, and API is the main de novo source of A5P in Gram-negative bacteria, because it is not readily available via glycolysis (16). API synthesizes A5P by catalyzing the reversible aldol-keto isomerization of D-ribulose 5-phosphate (Ru5P) to A5P (Fig. 1). It is reasonable to speculate that inhibition of API will have similar cellular effects as direct KDO synthase inhibition (7-9), thus providing another viable Gram-negative antibiotic target from the KDO pathway.The gene(s) encoding API in E. coli K-12 has yet to be positively identified. Recent work in this laboratory provided evidence that the open reading frame (ORF) yrbI encoded a 188-amino acid phosphatase that was highly specific for KDO 8-P (17). Analysis of the E. coli K-12 MG1655 genome (18) at the National Center for Biotechnology Information (NCBI) web site revealed an ORF (yrbH) located next to yrbI in the yrb cluster encoding a putative phosphosugar isomerase. The yrbH gene was cloned, overexpressed, and purified to homogeneity. Utilizing a newly developed 96-well microplate assay, the rec...

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Section: Discussionmentioning

confidence: 99%

Section: Identification Of Yrbh As Api In E Coli K-12-tomentioning

confidence: 99%

Escherichia coli YrbH Is a D-Arabinose 5-Phosphate Isomerase

Meredith

Woodard

2003

Journal of Biological Chemistry

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“…Establishing the requirement in E. coli is complicated by CMP-Kdo being essential for LPS biosynthesis and viability in wild-type bacteria (38), and by the duplication of genes encoding enzymes involved in CMP-Kdo biosynthesis in the kps locus. This limitation does not exist in N. meningitidis (37), where defects in the CMP-Kdo biosynthesis pathway lead to significant reductions in the amount of PSA CPS (37). However, it was not determined whether the residual CPS was lipidated, nor whether it was translocated to the cell surface.…”

Section: Discussionmentioning

confidence: 99%

“…Undecaprenyl monophospho-NeuAc has been proposed as the acceptor for the glycosyltransferases that synthesize the CPS glycan in the K1 system (39), invoking an elaborate two-step process involving transfer of such intermediates to a phospholipid (21,37). Presumably such a process would require a dedicated "ligase" enzyme, and there is no obvious candidate for this.…”

Section: Discussionmentioning

confidence: 99%

“…Several reports have suggested that Kdo biosynthesis plays a role in formation of CPS synthesized in the ABC transporterdependent pathway, but the molecular basis has not been established (21,33,37). Establishing the requirement in E. coli is complicated by CMP-Kdo being essential for LPS biosynthesis and viability in wild-type bacteria (38), and by the duplication of genes encoding enzymes involved in CMP-Kdo biosynthesis in the kps locus.…”

Section: Discussionmentioning

confidence: 99%

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Conserved glycolipid termini in capsular polysaccharides synthesized by ATP-binding cassette transporter-dependent pathways in Gram-negative pathogens

Willis

Stupak

Richards

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

117

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Bacterial capsules are surface layers made of long-chain polysaccharides. They are anchored to the outer membrane of many Gram-negative bacteria, including pathogens such as Escherichia coli , Neisseria meningitidis , Haemophilus influenzae , and Pasteurella multocida . Capsules protect pathogens from host defenses including complement-mediated killing and phagocytosis and therefore represent a major virulence factor. Capsular polysaccharides are synthesized by enzymes located in the inner (cytoplasmic) membrane and are then translocated to the cell surface. Whereas the enzymes that synthesize the polysaccharides have been studied in detail, the structure and biosynthesis of the anchoring elements have not been definitively resolved. Here we determine the structure of the glycolipid attached to the reducing terminus of the polysialic acid capsular polysaccharides from E. coli K1 and N. meningitidis group B and the heparosan-like capsular polysaccharide from E. coli K5. All possess the same unique glycolipid terminus consisting of a lyso-phosphatidylglycerol moiety with a β-linked poly-(3-deoxy- d - manno -oct-2-ulosonic acid) (poly-Kdo) linker attached to the reducing terminus of the capsular polysaccharide.

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Neisseria

Mietzner

Morse

2010

Topley &Amp; Wilson's Microbiology and Microbial Infections

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KpsF Is the Arabinose-5-phosphate Isomerase Required for 3-Deoxy-d-manno-octulosonic Acid Biosynthesis and for Both Lipooligosaccharide Assembly and Capsular Polysaccharide Expression in Neisseria meningitidis

Cited by 56 publications

References 58 publications

Escherichia coli YrbH Is a D-Arabinose 5-Phosphate Isomerase

Escherichia coli YrbH Is a D-Arabinose 5-Phosphate Isomerase

Conserved glycolipid termini in capsular polysaccharides synthesized by ATP-binding cassette transporter-dependent pathways in Gram-negative pathogens

Neisseria

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