Characterisation of a new cell wall teichoic acid produced by Listeria innocua ŽM39 and analysis of its biosynthesis genes

Bacillus anthracis, the causative agent of anthrax, replicates as chains of vegetative cells by regulating the separation of septal peptidoglycan. Surface (S)-layer proteins and associated proteins (BSLs) function as chain length determinants and bind to the secondary cell wall polysaccharide (SCWP). In this study, we identified theB. anthracislcpDmutant, which displays increased chain length and S-layer assembly defects due to diminished SCWP attachment to peptidoglycan. In contrast, theB. anthracislcpB3variant displayed reduced cell size and chain length, which could be attributed to increased deposition of BSLs. In other bacteria, LytR-CpsA-Psr (LCP) proteins attach wall teichoic acid (WTA) and polysaccharide capsule to peptidoglycan.B. anthracisdoes not synthesize these polymers, yet its genome encodes six LCP homologues, which, when expressed inS. aureus, promote WTA attachment. We propose a model wherebyB. anthracisLCPs promote attachment of SCWP precursors to discrete locations in the peptidoglycan, enabling BSL assembly and regulated separation of septal peptidoglycan.

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Control of teichoic acid and teichuronic acid biosynthesis in chemostat cultures of Bacillus subtilis var. niger

Biochemical Journal ◽

10.1042/bj1110001 ◽

1969 ◽

Vol 111 (1) ◽

pp. 1-5 ◽

Cited By ~ 76

Author(s):

D C Ellwood ◽

D. W. Tempest

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Teichoic Acid ◽

Extracellular Fluid ◽

Acid Synthesis ◽

Growth Cycle ◽

Wall Material ◽

Wall Teichoic Acid ◽

Anionic Polymers ◽

Teichuronic Acid

1. Quantitative determination of the anionic polymers present in the walls of Bacillus subtilis var. niger organisms undergoing transition, in a chemostat culture, from either Mg2+-limitation to PO43−-limitation or K+-limitation to PO43−-limitation showed that teichuronic acid synthesis started immediately the culture became PO43−-limited and proceeded at a rate substantially faster than the rate of biomass synthesis. 2. Simultaneously, the cell-wall teichoic acid content diminished at a rate greater than that due to dilution by newly synthesized wall material, and fragments of teichoic acid and mucopeptide accumulated in the culture extracellular fluid. 3. Equally rapid reverse changes occurred when a PO43−-limited B. subtilis var. niger culture was returned to being Mg2+-limited. 4. It is concluded that in this organism both teichoic acid and teichuronic acid syntheses are expressions of a single genotype, and a mechanism for the control of synthesis of both polymers is suggested. 5. These results are discussed with reference to the constantly changing environmental conditions that obtain in a batch culture and the variation in bacterial cell-wall composition that is reported to occur throughout the growth cycle.

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Genome-Wide Analysis of Phosphorylated PhoP Binding to Chromosomal DNA Reveals Several Novel Features of the PhoPR-Mediated Phosphate Limitation Response in Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.02570-14 ◽

2015 ◽

Vol 197 (8) ◽

pp. 1492-1506 ◽

Cited By ~ 14

Author(s):

Letal I. Salzberg ◽

Eric Botella ◽

Karsten Hokamp ◽

Haike Antelmann ◽

Sandra Maaß ◽

...

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Response Regulator ◽

Teichoic Acid ◽

Phosphate Limitation ◽

Chromosomal Dna ◽

Cell Wall Metabolism ◽

Wall Teichoic Acid ◽

Content Type ◽

Two Component

ABSTRACTThe PhoPR two-component signal transduction system controls one of three responses activated byBacillus subtilisto adapt to phosphate-limiting conditions (PHO response). The response involves the production of enzymes and transporters that scavenge for phosphate in the environment and assimilate it into the cell. However, inB. subtilisand some otherFirmicutesbacteria, cell wall metabolism is also part of the PHO response due to the high phosphate content of the teichoic acids attached either to peptidoglycan (wall teichoic acid) or to the cytoplasmic membrane (lipoteichoic acid). Prompted by our observation that the phosphorylated WalR (WalR∼P) response regulator binds to more chromosomal loci than are revealed by transcriptome analysis, we established the PhoP∼P bindome in phosphate-limited cells. Here, we show that PhoP∼P binds to the chromosome at 25 loci: 12 are within the promoters of previously identified PhoPR regulon genes, while 13 are newly identified. We extend the role of PhoPR in cell wall metabolism showing that PhoP∼P binds to the promoters of four cell wall-associated operons (ggaAB,yqgS,wapA, anddacA), although none show PhoPR-dependent expression under the conditions of this study. We also show that positive autoregulation ofphoPRexpression and full induction of the PHO response upon phosphate limitation require PhoP∼P binding to the 3′ end of thephoPRoperon.IMPORTANCEThe PhoPR two-component system controls one of three responses mounted byB. subtilisto adapt to phosphate limitation (PHO response). Here, establishment of the phosphorylated PhoP (PhoP∼P) bindome enhances our understanding of the PHO response in two important ways. First, PhoPR plays a more extensive role in adaptation to phosphate-limiting conditions than was deduced from transcriptome analyses. Among 13 newly identified binding sites, 4 are cell wall associated (ggaAB,yqgS,wapA, anddacA), revealing that PhoPR has an extended involvement in cell wall metabolism. Second, amplification of the PHO response must occur by a novel mechanism since positive autoregulation ofphoPRexpression requires PhoP∼P binding to the 3′ end of the operon.

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Structural features of cell wall teichoic acid and peptidoglycan of Actinomadura cremea INA 292

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1991.tb16126.x ◽

1991 ◽

Vol 199 (2) ◽

pp. 313-316 ◽

Cited By ~ 8

Author(s):

Natalia V. POTEKHINA ◽

Irina B. NAUMOVA ◽

Alexander S. SHASHKOV ◽

Larisa P. TEREKHOVA

Keyword(s):

Cell Wall ◽

Teichoic Acid ◽

Structural Features ◽

Wall Teichoic Acid

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Antagonism screen for inhibitors of bacterial cell wall biogenesis uncovers an inhibitor of undecaprenyl diphosphate synthase

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1511751112 ◽

2015 ◽

Vol 112 (35) ◽

pp. 11048-11053 ◽

Cited By ~ 44

Author(s):

Maya A. Farha ◽

Tomasz L. Czarny ◽

Cullen L. Myers ◽

Liam J. Worrall ◽

Shawn French ◽

...

Keyword(s):

Cell Wall ◽

High Throughput ◽

High Throughput Screening ◽

Teichoic Acid ◽

Antibacterial Drug ◽

Wall Teichoic Acid ◽

Chemical Screening ◽

Cellular Processes ◽

Cell Wall Biogenesis ◽

Antagonistic Interactions

Drug combinations are valuable tools for studying biological systems. Although much attention has been given to synergistic interactions in revealing connections between cellular processes, antagonistic interactions can also have tremendous value in elucidating genetic networks and mechanisms of drug action. Here, we exploit the power of antagonism in a high-throughput screen for molecules that suppress the activity of targocil, an inhibitor of the wall teichoic acid (WTA) flippase in Staphylococcus aureus. Well-characterized antagonism within the WTA biosynthetic pathway indicated that early steps would be sensitive to this screen; however, broader interactions with cell wall biogenesis components suggested that it might capture additional targets. A chemical screening effort using this approach identified clomiphene, a widely used fertility drug, as one such compound. Mechanistic characterization revealed the target was the undecaprenyl diphosphate synthase, an enzyme that catalyzes the synthesis of a polyisoprenoid essential for both peptidoglycan and WTA synthesis. The work sheds light on mechanisms contributing to the observed suppressive interactions of clomiphene and in turn reveals aspects of the biology that underlie cell wall synthesis in S. aureus. Further, this effort highlights the utility of antagonistic interactions both in high-throughput screening and in compound mode of action studies. Importantly, clomiphene represents a lead for antibacterial drug discovery.

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