An Escherichia coli undecaprenyl-pyrophosphate phosphatase implicated in undecaprenyl phosphate recycling

Tatar, Laura D.; Marolda, Cristina L.; Polischuk, Andrew N.; Leeuwen, Deborah van; Valvano, Miguel A.

doi:10.1099/mic.0.2007/006312-0

Cited by 94 publications

(117 citation statements)

References 45 publications

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“…Indeed, investigators who study WzxB function in Salmonella enterica do so in a genetic background where expression of O antigen is controlled by a galE mutation, which removes the first committed step in the biosynthetic pathway so that no Und-PP-linked O-antigen intermediates can accumulate until galactose is supplied (49,51,52). The deleterious effects of Und-P sequestration have also been noted for other transport mutants (53,54), as well as mutants that are unable to ligate O-antigen intermediates to the lipid A core (24) or that truncate the core oligosaccharide (55). In toto, then, our LPS and O-antigen findings are consistent with the conditional phenotypes that Und-P sequestration produces in strains with mutations that disrupt the synthesis of wall teichoic acids (56,57), capsular polysaccharides (58-60), exopolysaccharide (61), and ECA (30,62).…”

Section: Discussionmentioning

confidence: 88%

“…Mutations that prevent Und-P recycling lead to the accumulation of Und-PP-linked intermediates, resulting in a variety of deleterious or toxic effects (24)(25)(26)(27)(28). Because any one organism normally uses Und-P to synthesize multiple products, either these intermediates could be toxic in and of themselves or they might exert their effects indirectly by sequestering Und-P so that it becomes unavailable for use in other pathways.…”

mentioning

confidence: 99%

“…The resulting Und-P-linked oligosaccharides are translocated ("flipped") across the plasma membrane and transferred from the lipid carrier onto individual glycan chains (23). Either before or after being dephosphorylated, the leftover Und-PP carrier is then recycled into the cytoplasm, where it rejoins the pool of free Und-P (24).…”

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confidence: 99%

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Interrupting Biosynthesis of O Antigen or the Lipopolysaccharide Core Produces Morphological Defects in Escherichia coli by Sequestering Undecaprenyl Phosphate

Jorgenson

Young

2016

J Bacteriol

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Undecaprenyl phosphate (Und-P) is a member of the family of essential polyprenyl phosphate lipid carriers and in the Gramnegative bacterium Escherichia coli is required for synthesizing the peptidoglycan (PG) cell wall, enterobacterial common antigen (ECA), O antigen, and colanic acid. Previously, we found that interruption of ECA biosynthesis indirectly alters PG synthesis by sequestering Und-P via dead-end intermediates, causing morphological defects. To determine if competition for Und-P was a more general phenomenon, we determined if O-antigen intermediates caused similar effects. Indeed, disrupting the synthesis of O antigen or the lipopolysaccharide core oligosaccharide induced cell shape deformities, which were suppressed by preventing the initiation of O-antigen biosynthesis or by manipulating Und-P metabolism. We conclude that accumulation of O-antigen intermediates alters PG synthesis by sequestering Und-P. Importantly, many previous experiments addressed the physiological functions of various oligosaccharides and glycoconjugates, but these studies employed mutants that accumulate deleterious intermediates. Thus, conclusions based on these experiments must be reevaluated to account for possible indirect effects of Und-P sequestration. IMPORTANCEBacteria use long-chain isoprenoids like undecaprenyl phosphate (Und-P) as lipid carriers to assemble numerous glycan polymers that comprise the cell envelope. In any one bacterium, multiple oligosaccharide biosynthetic pathways compete for a common pool of Und-P, which means that disruptions in one pathway may produce secondary consequences that affect the others. Using the Gram-negative bacterium Escherichia coli as a model, we demonstrate that interruption of the biogenesis of O antigen, a major outer membrane component, indirectly impairs peptidoglycan synthesis by sequestering Und-P into dead-end intermediates. These results strongly argue that the functions of many Und-P-utilizing pathways must be reevaluated, because much of our current understanding is based on experiments that did not control for these unintended secondary effects. L ong-chain isoprenoids are widely conserved lipid carriers that translocate glycan intermediates across biological membranes, where they are assembled onto other polymers or released (reviewed in reference 1). In bacteria, the primary lipid carrier is undecaprenyl phosphate (Und-P), a 55-carbon isoprenoid (C 55 -P) also referred to as bactoprenol, which is the dephosphor ylated product of undecaprenyl pyrophosphate (Und-PP), as synthesized by UppS (2, 3). Homologues with shorter chains serve the same function in species of mycobacteria and corynebacteria (C 50 -P) (4, 5) and Paracoccus denitrificans (C 45 -P) (6). Und-P is required for synthesizing numerous bacterial glycans, including cell wall peptidoglycan (PG) (7,8), wall teichoic acids (WTA) (9), enterobacterial common antigen (ECA) (10), O antigen (11), capsular polysaccharides (12), several commercially important exopolysaccharides (13), and a variety of other...

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Section: Discussionmentioning

confidence: 88%

mentioning

confidence: 99%

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Interrupting Biosynthesis of O Antigen or the Lipopolysaccharide Core Produces Morphological Defects in Escherichia coli by Sequestering Undecaprenyl Phosphate

Jorgenson

Young

2016

J Bacteriol

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“…Here, we show that the PgpB active site faces the periplasm. Recently, Tatar et al (42) demonstrated that the signature residues of LpxT and YbjG were orientated toward the periplasmic space. However, the localization of the BacA active site remains still unknown.…”

Section: Discussionmentioning

confidence: 99%

Substrate Specificity and Membrane Topology of Escherichia coli PgpB, an Undecaprenyl Pyrophosphate Phosphatase

Touzé

Blanot

Mengin‐Lecreulx

2008

Journal of Biological Chemistry

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The synthesis of the lipid carrier undecaprenyl phosphate (C 55 -P) requires the dephosphorylation of its precursor, undecaprenyl pyrophosphate (C 55 -PP). The latter lipid is synthesized de novo in the cytosol and is also regenerated after its release from the C 55 -PP-linked glycans in the periplasm. In Escherichia coli the dephosphorylation of C 55 -PP was shown to involve four integral membrane proteins, BacA, and three members of the type 2 phosphatidic acid phosphatase family, PgpB, YbjG, and YeiU. Here, the PgpB protein was purified to homogeneity, and its phosphatase activity was examined. This enzyme was shown to catalyze the dephosphorylation of C 55 -PP with a relatively low efficiency compared with diacylglycerol pyrophosphate and farnesyl pyrophosphate (C 15 -PP) lipid substrates. However, the in vitro C 55 -PP phosphatase activity of PgpB was specifically enhanced by different phospholipids. We hypothesize that the phospholipids are important determinants to ensure proper conformation of the atypical long axis C 55 carrier lipid in membranes. Furthermore, a topological analysis demonstrated that PgpB contains six transmembrane segments, a large periplasmic loop, and the type 2 phosphatidic acid phosphatase signature residues at a periplasmic location.Undecaprenyl phosphate (C 55 -P) 2 is a 55-carbon-long polyprenol (see Fig. 1). It is an essential bacterial lipid required for the synthesis of various cell wall polymers such as peptidoglycan, lipopolysaccharides, teichoic acids, osmo-regulated periplasmic glucans, capsular polysaccharides, and the enterobacterial common antigen (1-10). C 55 -P is utilized as a carrier lipid that allows the transport of the hydrophilic oligosaccharide precursors across the cytoplasmic membrane toward the periplasm where the elongation of the glycan chains takes place. Accordingly, the precursor is linked to the carrier lipid via a pyrophosphate linkage (C 55 -PP-substrate) through the action of a specific glycosyltransferase at the cytosolic side of the inner membrane; thereafter, the complex is translocated through the membrane by a yet unknown mechanism, and finally, the glycosyl moiety is transferred to the appropriate expanding polymer. De novo synthesis of C 55 -P implicates two enzymatic steps (11, 12); it originates from undecaprenyl pyrophosphate (C 55 -PP), itself being synthesized by successive condensations of eight isopentenyl pyrophosphates (C 5 -PP) with farnesyl pyrophosphate (C 15 -PP) (Fig. 1) catalyzed by the cytosolic UppS enzyme, a cis-prenyl-pyrophosphate synthase (13, 14). The C 55 -PP must then be dephosphorylated to yield the active monophosphate form of the carrier lipid (11). C 55 -PP is not solely generated by de novo synthesis, but it is also released and recycled after the transfer of the oligosaccharide unit to the growing polymer in the periplasm. It is yet unclear on which side of the membrane C 55 -PP dephosphorylation occurs and how the carrier lipid is translocated across the membrane before being reused.The enzymatic dephosphoryl...

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“…105 Similarly, YbjG and YeiU signature residues were found to be oriented toward the periplasmic space. 102 It should be noted that the eukaryotic PAP2 homologue CWH8 was also found to be active on the extracytoplasmic side of the membrane, that is, the endoplasmic reticulum lumen, similar to the prokaryotic PAP2 enzymes. 27 In contrast, the membrane topology of the BacA protein, which lacks any feature of a typical phosphatase, remains to be explored.…”

Section: Recycling Versus De Novo Synthesismentioning

confidence: 99%

Deciphering the Metabolism of Undecaprenyl-Phosphate: The Bacterial Cell-Wall Unit Carrier at the Membrane Frontier

Manat

Roure

Auger

et al. 2014

Microbial Drug Resistance

120

139

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During the biogenesis of bacterial cell-wall polysaccharides, such as peptidoglycan, cytoplasmic synthesized precursors should be trafficked across the plasma membrane. This essential process requires a dedicated lipid, undecaprenyl-phosphate that is used as a glycan lipid carrier. The sugar is linked to the lipid carrier at the inner face of the membrane and is translocated toward the periplasm, where the glycan moiety is transferred to the growing polymer. Undecaprenyl-phosphate originates from the dephosphorylation of its precursor undecaprenyl-diphosphate, with itself generated by de novo synthesis or by recycling after the final glycan transfer. Undecaprenyl-diphosphate is de novo synthesized by the cytosolic cis-prenyltransferase undecaprenyldiphosphate synthase, which has been structurally and mechanistically characterized in great detail highlighting the condensation process. In contrast, the next step toward the formation of the lipid carrier, the dephosphorylation step, which has been overlooked for many years, has only started revealing surprising features. In contrast to the previous step, two unrelated families of integral membrane proteins exhibit undecaprenyldiphosphate phosphatase activity: BacA and members of the phosphatidic acid phosphatase type 2 super-family, raising the question of the significance of this multiplicity. Moreover, these enzymes establish an unexpected link between the synthesis of bacterial cell-wall polymers and other biological processes. In the present review, the current knowledge in the field of the bacterial lipid carrier, its mechanism of action, biogenesis, recycling, regulation, and future perspective works are presented.

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An Escherichia coli undecaprenyl-pyrophosphate phosphatase implicated in undecaprenyl phosphate recycling

Cited by 94 publications

References 45 publications

Interrupting Biosynthesis of O Antigen or the Lipopolysaccharide Core Produces Morphological Defects in Escherichia coli by Sequestering Undecaprenyl Phosphate

Interrupting Biosynthesis of O Antigen or the Lipopolysaccharide Core Produces Morphological Defects in Escherichia coli by Sequestering Undecaprenyl Phosphate

Substrate Specificity and Membrane Topology of Escherichia coli PgpB, an Undecaprenyl Pyrophosphate Phosphatase

Deciphering the Metabolism of Undecaprenyl-Phosphate: The Bacterial Cell-Wall Unit Carrier at the Membrane Frontier

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