Synthesis of CDP-Activated Ribitol for Teichoic Acid Precursors in Streptococcus pneumoniae

ABSTRACT Streptococcus pneumoniae has unusually complex cell wall teichoic acid and lipoteichoic acid, both of which contain a ribitol phosphate moiety. The lic region of the pneumococcal genome contains genes for the uptake and activation of choline, the attachment of phosphorylcholine to teichoic acid precursors, and the transport of these precursors across the cytoplasmic membrane. The role of two other, so far uncharacterized, genes, spr1148 and spr1149, in the lic region was determined. TarJ (spr1148) encodes an NADPH-dependent alcohol dehydrogenase for the synthesis of ribitol 5-phosphate from ribulose 5-phosphate. TarI (spr1149) encodes a cytidylyl transferase for the synthesis of cytidine 5′-diphosphate (CDP)-ribitol from ribitol 5-phosphate and cytidine 5′-triphosphate. We also present the crystal structure of TarI with and without bound CDP, and the structures present a rationale for the substrate specificity of this key enzyme. No transformants were obtained with insertion plasmids designed to interrupt the tarIJ genes, indicating that their function could be essential for cell growth. CDP-activated ribitol is a precursor for the synthesis of pneumococcal teichoic acids and some of the capsular polysaccharides. Thus, all eight genes in the lic region have a role in teichoic acid synthesis.

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Lipoteichoic acid of Streptococcus oralis Uo5: a novel biochemical structure comprising an unusual phosphorylcholine substitution pattern compared to Streptococcus pneumoniae

Scientific Reports ◽

10.1038/srep16718 ◽

2015 ◽

Vol 5 (1) ◽

Cited By ~ 15

Author(s):

Nicolas Gisch ◽

Dominik Schwudke ◽

Simone Thomsen ◽

Nathalie Heß ◽

Regine Hakenbeck ◽

...

Keyword(s):

Streptococcus Pneumoniae ◽

Teichoic Acid ◽

Structural Complexity ◽

Lipoteichoic Acid ◽

Close Relative ◽

Structural Variations ◽

Wall Teichoic Acid ◽

Substitution Pattern ◽

Type Iv ◽

Streptococcus Oralis

Abstract Members of the Mitis group of streptococci possess teichoic acids (TAs) as integral components of their cell wall that are unique among Gram-positive bacteria. Both, lipoteichoic (LTA) and wall teichoic acid, are formed by the same biosynthetic pathway, are of high complexity and contain phosphorylcholine (P-Cho) residues. These residues serve as anchors for choline-binding proteins (CBPs), some of which have been identified as virulence factors of the human pathogen Streptococcus pneumoniae. We investigated the LTA structure of its close relative Streptococcus oralis. Our analysis revealed that S. oralis Uo5 LTA has an overall architecture similar to pneumococcal LTA (pnLTA) and can be considered as a subtype of type IV LTA. Its structural complexity is even higher than that of pnLTA and its composition differs in number and type of carbohydrate moieties, inter-residue connectivities and especially the P-Cho substitution pattern. Here, we report the occurrence of a saccharide moiety substituted with two P-Cho residues, which is unique as yet in bacterial derived surface carbohydrates. Finally, we could link the observed important structural variations between S. oralis and S. pneumoniae LTA to the divergent enzymatic repertoire for their TA biosynthesis.

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LcpB Is a Pyrophosphatase Responsible for Wall Teichoic Acid Synthesis and Virulence in Staphylococcus aureus Clinical Isolate ST59

Frontiers in Microbiology ◽

10.3389/fmicb.2021.788500 ◽

2021 ◽

Vol 12 ◽

Author(s):

Ting Pan ◽

Jing Guan ◽

Yujie Li ◽

Baolin Sun

Keyword(s):

Staphylococcus Aureus ◽

Cell Wall ◽

Molecular Mechanisms ◽

Teichoic Acid ◽

Acid Synthesis ◽

Cell Wall Synthesis ◽

Wall Teichoic Acid ◽

Independent Manner ◽

Methicillin Resistant

The community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) causes severe pandemics primarily consisting of skin and soft tissue infections. However, the underlying pathomechanisms of the bacterium are yet to fully understood. The present study identifies LcpB protein, which belongs to the LytR-A-Psr (LCP) family, is crucial for cell wall synthesis and virulence in S. aureus. The findings revealed that LcpB is a pyrophosphatase responsible for wall teichoic acid synthesis. The results also showed that LcpB regulates enzyme activity through specific key arginine sites in its LCP domain. Furthermore, knockout of lcpB in the CA-MRSA isolate ST59 resulted in enhanced hemolytic activity, enlarged of abscesses, and increased leukocyte infiltration. Meanwhile, we also found that LcpB regulates virulence in agr-independent manner and the key sites for pyrophosphatase of LcpB play critical roles in regulating the virulence. In addition, the results showed that the role of LcpB was different between methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-sensitive Staphylococcus aureus (MSSA). This study therefore highlights the dual role of LcpB in cell wall synthesis and regulation of virulence. These insights on the underlying molecular mechanisms can thus guide the development of novel anti-infective strategies.

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d-Alanylation of Lipoteichoic Acid: Role of the d-Alanyl Carrier Protein in Acylation

Journal of Bacteriology ◽

10.1128/jb.183.6.2051-2058.2001 ◽

2001 ◽

Vol 183 (6) ◽

pp. 2051-2058 ◽

Cited By ~ 30

Author(s):

Michael Y. Kiriukhin ◽

Francis C. Neuhaus

Keyword(s):

Specific Interaction ◽

Teichoic Acid ◽

Lipoteichoic Acid ◽

Time Dependent ◽

Carrier Protein ◽

Wall Teichoic Acid ◽

Acyl Carrier Proteins ◽

Functional Specificity ◽

Hydrolysis Of

ABSTRACT The d-alanylation of membrane-associated lipoteichoic acid (LTA) in gram-positive organisms requires thed-alanine–d-alanyl carrier protein ligase (AMP) (Dcl) and the d-alanyl carrier protein (Dcp). Thedlt operon encoding these proteins (dltA anddltC) also includes dltB and dltD. dltB encodes a putative transport system, whiledltD encodes a protein which facilitates the binding of Dcp and Dcl for ligation with d-alanine and has thioesterase activity for mischarged d-alanyl-acyl carrier proteins (ACPs). In previous results it was shown that d-alanyl-Dcp donates its ester residue to membrane-associated LTA (M. P. Heaton and F. C. Neuhaus, J. Bacteriol. 176: 681–690, 1994). However, all efforts to identify an enzyme which catalyzes thisd-alanylation process were unsuccessful. It was discovered that incubation of d-alanyl-Dcp in the presence of LTA resulted in the time-dependent hydrolysis of this d-alanyl thioester. d-Alanyl-ACP in the presence of LTA was not hydrolyzed. When Dcp was incubated with membrane-associatedd-alanyl LTA, a time and concentration-dependent formation of d-alanyl-Dcp was found. The addition of NaCl to this reaction inhibited the formation of d-alanyl-Dcp and stimulated the hydrolysis of d-alanyl-Dcp. Since these reactions are specific for the carrier protein (Dcp), it is suggested that Dcp has a unique binding site which interacts with the poly(Gro-P) moiety of LTA. It is this specific interaction that provides the functional specificity for the d-alanylation process. The reversibility of this process provides a mechanism for the transacylation of the d-alanyl ester residues between LTA and wall teichoic acid.

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Mutants of Bacillus subtilis 168 Thermosensitive for Growth and Wall Teichoic Acid Synthesis

Microbiology ◽

10.1099/00221287-135-5-1325 ◽

1989 ◽

Vol 135 (5) ◽

pp. 1325-1334 ◽

Cited By ~ 6

Author(s):

M. Briehl ◽

H. M. Pooley ◽

D. Karamata

Keyword(s):

Bacillus Subtilis ◽

Teichoic Acid ◽

Acid Synthesis ◽

Wall Teichoic Acid ◽

Bacillus Subtilis 168

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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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Taking aim at wall teichoic acid synthesis: new biology and new leads for antibiotics

The Journal of Antibiotics ◽

10.1038/ja.2013.100 ◽

2013 ◽

Vol 67 (1) ◽

pp. 43-51 ◽

Cited By ~ 60

Author(s):

Edward WC Sewell ◽

Eric D Brown

Keyword(s):

Teichoic Acid ◽

Acid Synthesis ◽

Wall Teichoic Acid ◽

New Biology

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Analysis of Bacillus subtilis tagAB andtagDEF Expression during Phosphate Starvation Identifies a Repressor Role for PhoP∼P

Journal of Bacteriology ◽

10.1128/jb.180.3.753-758.1998 ◽

1998 ◽

Vol 180 (3) ◽

pp. 753-758 ◽

Cited By ~ 58

Author(s):

Wei Liu ◽

Stephen Eder ◽

F. Marion Hulett

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Teichoic Acid ◽

Phosphate Starvation ◽

Acid Synthesis ◽

Pho Regulon ◽

Wall Teichoic Acid ◽

Negative Role ◽

First Time ◽

Starvation Conditions

ABSTRACT The tagAB and tagDEF operons, which are adjacent and divergently transcribed, encode genes responsible for cell wall teichoic acid synthesis in Bacillus subtilis. TheBacillus data presented here suggest that PhoP and PhoR are required for direct repression of transcription of the two operons under phosphate starvation conditions but have no regulatory role under phosphate-replete conditions. These data identify for the first time that PhoP∼P has a negative role in Pho regulon gene regulation.

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Role of PhoP∼P in Transcriptional Regulation of Genes Involved in Cell Wall Anionic Polymer Biosynthesis in Bacillus subtilis

Journal of Bacteriology ◽

10.1128/jb.180.15.4007-4010.1998 ◽

1998 ◽

Vol 180 (15) ◽

pp. 4007-4010 ◽

Cited By ~ 29

Author(s):

Ying Qi ◽

F. Marion Hulett

Keyword(s):

Cell Wall ◽

Bacillus Subtilis ◽

Teichoic Acid ◽

Acid Synthesis ◽

In Vitro Transcription ◽

Anionic Polymer ◽

Transcription System ◽

Teichuronic Acid

ABSTRACT tagA, tagD, and tuaA operons are responsible for the synthesis of cell wall anionic polymer, teichoic acid, and teichuronic acid, respectively, in Bacillus subtilis. Under phosphate starvation conditions, teichuronic acid is synthesized while teichoic acid synthesis is inhibited. Expression of these genes is controlled by PhoP-PhoR, a two-component system. It has been proposed that PhoP∼P plays a key role in the activation oftuaA and the repression of tagA andtagD. In this study, we demonstrated the role of PhoP∼P in the switch process from teichoic acid synthesis to teichuronic acid synthesis, by using an in vitro transcription system. The results indicate that PhoP∼P is sufficient to repress the transcription of the tagA and tagD promoters and also to activate the transcription of the tuaA promoter.

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