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

2015 ◽  
Vol 5 (1) ◽  
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.

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

2008 ◽  
Vol 191 (4) ◽  
pp. 1200-1210 ◽  
Author(s):  
Stefanie Baur ◽  
Jon Marles-Wright ◽  
Stephan Buckenmaier ◽  
Richard J. Lewis ◽  
Waldemar Vollmer
Keyword(s):  
Streptococcus Pneumoniae ◽  
Cytoplasmic Membrane ◽  
Teichoic Acid ◽  
Lipoteichoic Acid ◽  
Acid Synthesis ◽  
Capsular Polysaccharides ◽  
Complex Cell ◽  
Wall Teichoic Acid ◽  
Key Enzyme

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.

scholarly journals Analysis of phage-resistant mechanisms inStaphylococcus aureusSA003 reveals a different binding mechanism for the closely related Twort-like phages ϕSA012 and ϕSA039

2018 ◽  
Author(s):  
Aa Haeruman Azam ◽  
Fumiya Hoshiga ◽  
Ippei Takeuchi ◽  
Kazuhiko Miyanaga ◽  
Yasunori Tanji
Keyword(s):  
Teichoic Acid ◽  
Point Mutations ◽  
In Silico Analysis ◽  
Close Relative ◽  
Phenotypic Change ◽  
Missense Mutations ◽  
Wild Type ◽  
Wall Teichoic Acid ◽  
Whole Genome Analysis ◽  
Specific Phage

ABSTRACTWe have previously generated strains ofStaphylococcus aureusSA003 resistant to its specific phage ϕSA012 through long-term coevolution experiment. However, the DNA mutations responsible for the phenotypic change of phage resistance are unknown. Whole-genome analysis revealed six genes that acquired unique point mutations: five missense mutations and one nonsense mutation. Moreover, one deletion, 1.779-bp, resulted in the deletion of the genes encoding glycosyltransferase, TarS, and iron-sulfure repair protein, ScdA. The deletion occurred from the second round of coculture (SA003R2) and remained through the last round. The ϕSA012 infection toward SA003R2 had decreased to 79.77±7.50% according to plating efficiency. Complementation of the phage-resistant strain by the wild-type allele showed two mutated host genes were linked to the inhibition of post-adsorption, and five genes were linked to phage adsorption of ϕSA012. Unlike ϕSA012, infection by ϕSA039, a close relative of ϕSA012, onto SA003R2 was impaired drastically. Complementation of SA003R2 by wild-typetarSrestores the infectivity of ϕSA039. Thus, we concluded that ϕSA039 requires β-GlcNAc in Wall Teichoic Acid (WTA) for its binding. In silico analysis of the ϕSA039 genome revealed that several proteins in the tail and baseplate region were different from ϕSA012; notably the partial deletion oforf96of ϕSA039, a homolog oforf99of ϕSA012.Orf100of ϕSA039, a homolog ofOrf103of ϕSA012, a previously reported receptor binding protein (RBP), had low similarity (86%) to that of ϕSA012. The difference in tail and baseplate proteins might be the factor for specificity difference between ϕSA012 and ϕSA039.

scholarly journals Mutations in the tacF Gene of Clinical Strains and Laboratory Transformants of Streptococcus pneumoniae: Impact on Choline Auxotrophy and Growth Rate

2008 ◽  
Vol 190 (12) ◽  
pp. 4129-4138 ◽  
Author(s):  
Ana González ◽  
Daniel Llull ◽  
María Morales ◽  
Pedro García ◽  
Ernesto García
Keyword(s):  
Streptococcus Pneumoniae ◽  
Repeat Unit ◽  
Teichoic Acid ◽  
Integral Membrane Protein ◽  
Nutritional Requirement ◽  
Spontaneous Mutant ◽  
Free Medium ◽  
Streptococcus Oralis ◽  
Complex Relationships ◽  
Lipoteichoic Acids

ABSTRACT The nutritional requirement that Streptococcus pneumoniae has for the aminoalcohol choline as a component of teichoic and lipoteichoic acids appears to be exclusive to this prokaryote. A mutation in the tacF gene, which putatively encodes an integral membrane protein (possibly, a teichoic acid repeat unit transporter), has been recently identified as responsible for generating a choline-independent phenotype of S. pneumoniae (M. Damjanovic, A. S. Kharat, A. Eberhardt, A. Tomasz, and W. Vollmer, J. Bacteriol. 189:7105-7111, 2007). We now report that Streptococcus mitis can grow in choline-free medium, as previously illustrated for Streptococcus oralis. While we confirmed the finding by Damjanovic et al. of the involvement of TacF in the choline dependence of the pneumococcus, the genetic transformation of S. pneumoniae R6 by using S. mitis SK598 DNA and several PCR-amplified tacF fragments suggested that a minimum of two mutations were required to confer improved fitness to choline-independent S. pneumoniae mutants. This conclusion is supported by sequencing results also reported here that indicate that a spontaneous mutant of S. pneumoniae (strain JY2190) able to proliferate in the absence of choline (or analogs) is also a double mutant for the tacF gene. Microscopic observations and competition experiments during the cocultivation of choline-independent strains confirmed that a minimum of two amino acid changes were required to confer improved fitness to choline-independent pneumococcal strains when growing in medium lacking any aminoalcohol. Our results suggest complex relationships among the different regions of the TacF teichoic acid repeat unit transporter.

scholarly journals Inactivation of the monofunctional peptidoglycan glycosyltransferase SgtB allowsStaphylococcus aureusto survive in the absence of lipoteichoic acid

2018 ◽  
Author(s):  
Eleni Karinou ◽  
Christopher F. Schuster ◽  
Manuel Pazos ◽  
Waldemar Vollmer ◽  
Angelika Gründling
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Wall ◽  
Teichoic Acid ◽  
Fold Increase ◽  
Lipoteichoic Acid ◽  
Wall Teichoic Acid ◽  
Anionic Polymers ◽  
Peptidoglycan Structure ◽  
Intimate Link ◽  
Inactivating Mutations

AbstractThe cell wall ofStaphylococcus aureusis composed of peptidoglycan and the anionic polymers lipoteichoic acid (LTA) and wall teichoic acid. LTA is required for growth and normal cell morphology inS. aureus.Strains lacking LTA are usually only viable when grown under osmotically stabilizing conditions or after the acquisition of compensatory mutations. LTA negative suppressor strains with inactivating mutations ingdpP, resulting in an increase in intracellular c-di-AMP levels, have been described previously. Here, we sought to identify factors other than c-di-AMP that allowS. aureusto survive without LTA. LTA-negative strains able to grow in un-supplemented medium were obtained and found to contain mutations insgtB, mazE, clpXorvraT. The growth improvement through mutations inmazEandsgtBwas confirmed by complementation analysis. We also show that anS. aureus sgtBtransposon mutant, inactivated for the monofunctional peptidoglycan glycosyltransferase SgtB, displays a 4-fold increase in the MIC towards a number of cell wall-targeting antibiotics, suggesting that alteration in the peptidoglycan structure could help bacteria compensate for the lack of LTA. Muropeptide analysis of peptidoglycan isolated from a WT andsgtBmutant strains did not reveal any sizable alternations in the peptidoglycan structure. In contrast, the peptidoglycan isolated from an LTA-negativeltaSmutant strain showed a significant reduction in the fraction of highly crosslinked peptidoglycan, which was partially rescued in thesgtB/ltaSdouble mutant suppressor strain. Taken together, these data point towards an important function of LTA in cell wall integrity through its requirement for proper peptidoglycan assembly.ImportanceThe bacterial cell wall acts as primary defence against environmental insults such as changes in osmolarity. It is also a vulnerable structure as defects in its synthesis can lead to growth arrest or cell death. The important human pathogenStaphylococcus aureushas a typical Gram-positive cell wall, which consists of peptidoglycan and the anionic polymers lipoteichoic acid (LTA) and wall teichoic acid. Several clinically relevant antibiotics inhibit the synthesis of peptidoglycan; hence it and teichoic acids are considered attractive targets for the development of new antimicrobials. We show that LTA is required for efficient peptidoglycan crosslinking inS. aureusand inactivation of a peptidoglycan glycosyltransferase can partially rescue this defect, altogether revealing an intimate link between peptidoglycan and LTA synthesis.

scholarly journals Incorporation of D-Alanine into Lipoteichoic Acid and Wall Teichoic Acid inBacillus subtilis

1995 ◽  
Vol 270 (26) ◽  
pp. 15598-15606 ◽  
Author(s):  
Marta Perego ◽  
Philippe Glaser ◽  
Antonia Minutello ◽  
Mark A. Strauch ◽  
Klaus Leopold ◽  
...  
Keyword(s):  
Teichoic Acid ◽  
Lipoteichoic Acid ◽  
Wall Teichoic Acid

scholarly journals Generation and Properties of a Streptococcus pneumoniae Mutant Which Does Not Require Choline or Analogs for Growth

1998 ◽  
Vol 180 (8) ◽  
pp. 2093-2101 ◽  
Author(s):  
Janet Yother ◽  
Klaus Leopold ◽  
Johanna White ◽  
Werner Fischer
Keyword(s):  
Streptococcus Pneumoniae ◽  
Stationary Phase ◽  
Cell Walls ◽  
Parent Strain ◽  
Protein A ◽  
Teichoic Acid ◽  
Surface Protein ◽  
Chain Structure ◽  
Wall Teichoic Acid ◽  
Mutant Cells

A mutant (JY2190) of Streptococcus pneumoniae Rx1 which had acquired the ability to grow in the absence of choline and analogs was isolated. Lipoteichoic acid (LTA) and wall teichoic acid (TA) isolated from the mutant were free of phosphocholine and other phosphorylated amino alcohols. Both polymers showed an unaltered chain structure and, in the case of LTA, an unchanged glycolipid anchor. The cell wall composition was also not altered except that, due to the lack of phosphocholine, the phosphate content of cell walls was half that of the parent strain. Isolated cell walls of the mutant were resistant to hydrolysis by pneumococcal autolysin (N-acetylmuramyl-l-alanine amidase) but were cleaved by the muramidases CPL and cellosyl. The lack of active autolysin in the mutant cells became apparent by impaired cell separation at the end of cell division and by resistance against stationary-phase and penicillin-induced lysis. As a result of the absence of choline in the LTA, pneumococcal surface protein A (PspA) was no longer retained on the cytoplasmic membrane. During growth in the presence of choline, which was incorporated as phosphocholine into LTA and TA, the mutant cells separated normally, did not release PspA, and became penicillin sensitive. However, even under these conditions, they did not lyse in the stationary phase, and they showed poor reactivity with antibody to phosphocholine and an increased release of C-polysaccharide from the cell. In contrast to ethanolamine-grown parent cells (A. Tomasz, Proc. Natl. Acad. Sci. USA 59:86–93, 1968), the choline-free mutant cells retained the capability to undergo genetic transformation but, compared to Rx1, with lower frequency and at an earlier stage of growth. The properties of the mutant could be transferred to the parent strain by DNA of the mutant.

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

2001 ◽  
Vol 183 (6) ◽  
pp. 2051-2058 ◽  
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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