The envA Permeability/Cell Division Gene of Escherichia coli Encodes the Second Enzyme of Lipid A Biosynthesis

Young, Katherine; Silver, Lynn L.; Bramhill, David; Cameron, Patricia M.; Eveland, Suzanne S.; Raetz, Christian R. H.; Hyland, Sheryl A.; Anderson, Matt S.

doi:10.1074/jbc.270.51.30384

Cited by 180 publications

(158 citation statements)

References 39 publications

(40 reference statements)

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“…Based on the ability of OME to rescue motility and development in mutants with truncated LPS, we sought to create a model of membrane damage that interrupts the biosynthesis of the whole LPS molecule and thus affects cell viability. Lipid A is the acylated disaccharide that embeds LPS in the OM and is essential in most Gram-negative bacteria (27). The first committed step in the biosynthesis of lipid A is the deacetylation of UDP-3-O-acyl N-acetyl-glucosamine by cytoplasmically localized LpxC (27), a protein that is highly conserved and essential across Gram-negative bacterial species (28).…”

Section: Ome Rescues the Motility And Development Of O-antigen Mutantsmentioning

confidence: 99%

“…Lipid A is the acylated disaccharide that embeds LPS in the OM and is essential in most Gram-negative bacteria (27). The first committed step in the biosynthesis of lipid A is the deacetylation of UDP-3-O-acyl N-acetyl-glucosamine by cytoplasmically localized LpxC (27), a protein that is highly conserved and essential across Gram-negative bacterial species (28). We thus constructed a strain of M. xanthus in which the endogenous lpxC promoter was replaced with a vanillate-inducible promoter (29) (P van ::lpxC) (Fig.…”

Section: Ome Rescues the Motility And Development Of O-antigen Mutantsmentioning

confidence: 99%

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Cell rejuvenation and social behaviors promoted by LPS exchange in myxobacteria

Vassallo

Pathak

Cao

et al. 2015

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

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Bacterial cells in their native environments must cope with factors that compromise the integrity of the cell. The mechanisms of coping with damage in a social or multicellular context are poorly understood. Here we investigated how a model social bacterium, Myxococcus xanthus, approaches this problem. We focused on the social behavior of outer membrane exchange (OME), in which cells transiently fuse and exchange their outer membrane (OM) contents. This behavior requires TraA, a homophilic cell surface receptor that identifies kin based on similarities in a polymorphic region, and the TraB cohort protein. As observed by electron microscopy, TraAB overexpression catalyzed a prefusion OM junction between cells. We then showed that damage sustained by the OM of one population was repaired by OME with a healthy population. Specifically, LPS mutants that were defective in motility and sporulation were rescued by OME with healthy donors. In addition, a mutant with a conditional lethal mutation in lpxC, an essential gene required for lipid A biosynthesis, was rescued by Tradependent interactions with a healthy population. Furthermore, lpxC cells with damaged OMs, which were more susceptible to antibiotics, had resistance conferred to them by OME with healthy donors. We also show that OME has beneficial fitness consequences to all cells. Here, in merged populations of damaged and healthy cells, OME catalyzed a dilution of OM damage, increasing developmental sporulation outcomes of the combined population by allowing it to reach a threshold density. We propose that OME is a mechanism that myxobacteria use to overcome cell damage and to transition to a multicellular organism.Myxococcus xanthus | outer membrane | lipopolysaccharide | lpxC | fusion A fundamental question in biology is how cells cope with damage. Microbes occupy diverse habitats fraught with physical, biological, and chemical insults (1, 2). UV radiation, desiccation, predation, extracellular enzymes, antimicrobial compounds, pH, temperature, and osmolarity changes are all stresses to the individual cell. In addition, when cells are in nutrient-poor environments, cell division can be rare, taking days to months to complete (3). In a slow-growing state, cell-surface components that may not be undergoing active repair can accumulate damage through natural aging processes such as oxidation (4, 5) and protein denaturation. Although internal cell stress response pathways are known (6), mechanisms to cope with cell surface damage are less well understood.Although cell damage threatens the fitness of the individual, social organisms have strength in numbers. The strategy of kin selection allows evolutionarily viable cooperation between individuals in a closely related population (7). Communication between individuals and sharing of resources establishes the potential for assistance between individual population members. Social support can be beneficial when the fitness of individuals in a group depends on collaborative behaviors such as prey hunting or the developm...

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Section: Ome Rescues the Motility And Development Of O-antigen Mutantsmentioning

confidence: 99%

Section: Ome Rescues the Motility And Development Of O-antigen Mutantsmentioning

confidence: 99%

Cell rejuvenation and social behaviors promoted by LPS exchange in myxobacteria

Vassallo

Pathak

Cao

et al. 2015

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

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“…The first committed step in lipid A biosynthesis is catalyzed by a metal-dependent deacetylase, UDP-(3-O-((R)-3-hydroxymyristoyl))-N-acetylglucosamine deacetylase (LpxC), which hydrolyzes UDP-(3-O-((R)-3-hydroxymyristoyl))-N-acetylglucosamine to form acetate and UDP-(3-O-((R)-3-hydroxymyristoyl))glucosamine (11)(12)(13)(14). The three-dimensional structure of LpxC from Aquifex aeolicus has been determined by X-ray crystallography (15) and NMR spectroscopy (16,17) and reveals a novel α + β fold and the new zinc-binding motif, HKX(L,F) D. In the mechanism first proposed based on analysis of the X-ray crystal structure (15), E78 serves as a general base by abstracting a proton from the zinc-bound water molecule to promote nucleophilic attack at the substrate, and this aspect of the catalytic mechanism has been probed in detailed structural and enzymological studies (17)(18)(19) Figure 1) (19,20).…”

Section: Nih Public Accessmentioning

confidence: 99%

Mechanistic Inferences from the Binding of Ligands to LpxC, a Metal-Dependent Deacetylase^,

Gennadios

Whittington

et al. 2006

Biochemistry

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The metal-dependent deacetylase LpxC catalyzes the first committed step of lipid A biosynthesis in Gram-negative bacteria. Accordingly, LpxC is an attractive target for the development of inhibitors that may serve as potential new antibiotics for the treatment of Gram-negative bacterial infections. Here, we report the 2.7 Å resolution X-ray crystal structure of LpxC complexed with the substrate analogue inhibitor, TU-514, and the 2.0 Å resolution structure of LpxC complexed with imidazole. The X-ray crystal structure of LpxC complexed with TU-514 allows for a detailed examination of the coordination geometry of the catalytic zinc ion and other enzyme-inhibitor interactions in the active site. The hydroxamate group of TU-514 forms a bidentate chelate complex with the zinc ion and makes hydrogen bond interactions with conserved active site residues E78, H265, and T191. The inhibitor C-4 hydroxyl group makes direct hydrogen bond interactions with E197 and H58. Finally, the C-3 myristate moiety of the inhibitor binds in the hydrophobic tunnel of the active site. These intermolecular interactions provide a foundation for understanding structural aspects of enzyme-substrate and enzyme-inhibitor affinity. Comparison of the TU-514 complex with cacodylate and imidazole complexes suggests a possible substrate diphosphate binding site and highlights residues that may stabilize the tetrahedral intermediate and its flanking transition states in catalysis. Evidence is also presented for a catalytic zinc ion in the native zinc enzyme coordinated by H79, H238, D242, and two water molecules with square pyramidal geometry. These results suggest that the native state of this metallohydrolase may contain a pentacoordinate zinc ion, which contrasts with the native states of archetypical zinc hydrolases such as thermolysin and carboxypeptidase A.The outer leaflet of the outer membrane of a Gram-negative bacterium is composed of lipopolysaccharide (LPS), which serves as a permeability barrier that protects the bacterium against erythromycin and other antibiotics (1-6). 1 Each LPS molecule contains O-antigen † This work was supported by National Institutes of Health grants GM49758 (D. W. C.) and GM40602 (C. A. F.) ‡ The atomic coordinates for LpxC complexed with TU-514 and imidazole have been deposited in the Protein Data Bank, www.rcsb.org, with accession codes 2GO4 and 2GO3, respectively. * To whom correspondence should be addressed. Telephone: (215) 898-5714. Fax: (215) 573-2201. Email: chris@sas.upenn.edu. § University of Pennsylvania ⊥ Current address: Amgen, Inc., One Kendall Square, Bldg. 1000, Cambridge, MA 02139 ‖ University of Alberta; current address: Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford St., Cambridge, MA 02138 # University of Michigan 1 Abbreviations: LPS, lipopolysaccharide; TU-514, 1,5-anhydro-2-C-(carboxymethyl-N-hydroxamide)-2-deoxy-3-O-myristoyl-Dglucitol; UDP, uridine 5′-diphosphate; NCS, noncrystallographic symmetry; HEPES, 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic ...

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“…1) (8). Because LpxA catalyzes a thermodynamically unfavorable reaction (K eq Ϸ 0.01) (8), the second enzyme of the pathway, the deacetylase (LpxC), is the committed step (9,10). LpxA and LpxC are essential for growth (10)(11)(12), and both are validated targets for the design of novel antibiotics (13)(14)(15).…”

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confidence: 99%

Structural basis for the acyl chain selectivity and mechanism of UDP- N -acetylglucosamine acyltransferase

Williams

Raetz

2007

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

146

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The envA Permeability/Cell Division Gene of Escherichia coli Encodes the Second Enzyme of Lipid A Biosynthesis

Cited by 180 publications

References 39 publications

Cell rejuvenation and social behaviors promoted by LPS exchange in myxobacteria

Cell rejuvenation and social behaviors promoted by LPS exchange in myxobacteria

Mechanistic Inferences from the Binding of Ligands to LpxC, a Metal-Dependent Deacetylase^,

Structural basis for the acyl chain selectivity and mechanism of UDP- N -acetylglucosamine acyltransferase

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The envA Permeability/Cell Division Gene of Escherichia coli Encodes the Second Enzyme of Lipid A Biosynthesis

Cited by 180 publications

References 39 publications

Cell rejuvenation and social behaviors promoted by LPS exchange in myxobacteria

Cell rejuvenation and social behaviors promoted by LPS exchange in myxobacteria

Mechanistic Inferences from the Binding of Ligands to LpxC, a Metal-Dependent Deacetylase,

Structural basis for the acyl chain selectivity and mechanism of UDP- N -acetylglucosamine acyltransferase

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Mechanistic Inferences from the Binding of Ligands to LpxC, a Metal-Dependent Deacetylase^,