4,6-O-Pyruvyl Ketal Modified N-Acetylmannosamine of the Secondary Cell Wall Polysaccharide of Bacillus anthracis Is the Anchoring Residue for Its Surface Layer Proteins

2018 ◽  
Vol 140 (49) ◽  
pp. 17079-17085 ◽  
Author(s):  
Robert N. Chapman ◽  
Lin Liu ◽  
Geert-Jan Boons
Keyword(s):  
Cell Wall ◽  
Surface Layer ◽  
Bacillus Anthracis ◽  
Secondary Cell Wall ◽  
Cell Wall Polysaccharide ◽  
Surface Layer Proteins

scholarly journals LytR-CpsA-Psr Enzymes as Determinants of Bacillus anthracis Secondary Cell Wall Polysaccharide Assembly

2014 ◽  
Vol 197 (2) ◽  
pp. 343-353 ◽  
Author(s):  
Megan Liszewski Zilla ◽  
Yvonne G. Y. Chan ◽  
Justin Mark Lunderberg ◽  
Olaf Schneewind ◽  
Dominique Missiakas
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Chain Length ◽  
Secondary Cell Wall ◽  
Teichoic Acid ◽  
Cell Wall Polysaccharide ◽  
Wall Teichoic Acid ◽  
Content Type ◽  
Associated Proteins ◽  
Layer Assembly

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.

scholarly journals Genes Required for Bacillus anthracis Secondary Cell Wall Polysaccharide Synthesis

2016 ◽  
Vol 199 (1) ◽  
Author(s):  
So-Young Oh ◽  
J. Mark Lunderberg ◽  
Alice Chateau ◽  
Olaf Schneewind ◽  
Dominique Missiakas
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Vegetative Growth ◽  
Cell Shape ◽  
Secondary Cell Wall ◽  
Cell Wall Polysaccharide ◽  
Genetic Determinants ◽  
Content Type ◽  
Repeat Structure ◽  
Layer Assembly

ABSTRACT The secondary cell wall polysaccharide (SCWP) is thought to be essential for vegetative growth and surface (S)-layer assembly in Bacillus anthracis; however, the genetic determinants for the assembly of its trisaccharide repeat structure are not known. Here, we report that WpaA (BAS0847) and WpaB (BAS5274) share features with membrane proteins involved in the assembly of O-antigen lipopolysaccharide in Gram-negative bacteria and propose that WpaA and WpaB contribute to the assembly of the SCWP in B. anthracis. Vegetative forms of the B. anthracis wpaA mutant displayed increased lengths of cell chains, a cell separation defect that was attributed to mislocalization of the S-layer-associated murein hydrolases BslO, BslS, and BslT. The wpaB mutant was defective in vegetative replication during early logarithmic growth and formed smaller colonies. Deletion of both genes, wpaA and wpaB, did not yield viable bacilli, and when depleted of both wpaA and wpaB, B. anthracis could not maintain cell shape, support vegetative growth, or assemble SCWP. We propose that WpaA and WpaB fulfill overlapping glycosyltransferase functions of either polymerizing repeat units or transferring SCWP polymers to linkage units prior to LCP-mediated anchoring of the polysaccharide to peptidoglycan. IMPORTANCE The secondary cell wall polysaccharide (SCWP) is essential for Bacillus anthracis growth, cell shape, and division. SCWP is comprised of trisaccharide repeats (→4)-β-ManNAc-(1→4)-β-GlcNAc-(1→6)-α-GlcNAc-(1→) with α-Gal and β-Gal substitutions; however, the genetic determinants and enzymes for SCWP synthesis are not known. Here, we identify WpaA and WpaB and report that depletion of these factors affects vegetative growth, cell shape, and S-layer assembly. We hypothesize that WpaA and WpaB are involved in the assembly of SCWP prior to transfer of this polymer onto peptidoglycan.

scholarly journals Bacillus anthracistagOIs Required for Vegetative Growth and Secondary Cell Wall Polysaccharide Synthesis

2015 ◽  
Vol 197 (22) ◽  
pp. 3511-3520 ◽  
Author(s):  
J. Mark Lunderberg ◽  
Megan Liszewski Zilla ◽  
Dominique Missiakas ◽  
Olaf Schneewind
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Vegetative Growth ◽  
Cell Shape ◽  
Secondary Cell Wall ◽  
Cell Wall Polysaccharide ◽  
Content Type ◽  
Phosphodiester Bonds ◽  
Acid Labile ◽  
Layer Assembly

ABSTRACTBacillus anthraciselaborates a linear secondary cell wall polysaccharide (SCWP) that retains surface (S)-layer and associated proteins via their S-layer homology (SLH) domains. The SCWP is comprised of trisaccharide repeats [→4)-β-ManNAc-(1→4)-β-GlcNAc-(1→6)-α-GlcNAc-(1→] and tethered via acid-labile phosphodiester bonds to peptidoglycan. Earlier work identified UDP-GlcNAc 2-epimerases GneY (BAS5048) and GneZ (BAS5117), which act as catalysts of ManNAc synthesis, as well as a polysaccharide deacetylase (BAS5051), as factors contributing to SCWP synthesis. Here, we show thattagO(BAS5050), which encodes a UDP-N-acetylglucosamine:undecaprenyl-PN-acetylglucosaminyl 1-P transferase, the enzyme that initiates the synthesis of murein linkage units, is required forB. anthracisSCWP synthesis and S-layer assembly. Similar togneY-gneZmutants,B. anthracisstrains lackingtagOcannot maintain cell shape or support vegetative growth. In contrast, mutations in BAS5051 do not affectB. anthraciscell shape, vegetative growth, SCWP synthesis, or S-layer assembly. These data suggest that TagO-mediated murein linkage unit assembly supports SCWP synthesis and attachment to the peptidoglycan via acid-labile phosphodiester bonds. Further,B. anthracisvariants unable to synthesize SCWP trisaccharide repeats cannot sustain cell shape and vegetative growth.IMPORTANCEBacillus anthraciselaborates an SCWP to support vegetative growth and envelope assembly. Here, we show that some, but not all, SCWP synthesis is dependent ontagO-derived murein linkage units and subsequent attachment of SCWP to peptidoglycan. The data implicate secondary polymer modifications of peptidoglycan and subcellular distributions as a key feature of the cell cycle in Gram-positive bacteria and establish foundations for work on the molecular functions of the SCWP and on inhibitors with antibiotic attributes.

scholarly journals Bacillus anthracis Acetyltransferases PatA1 and PatA2 Modify the Secondary Cell Wall Polysaccharide and Affect the Assembly of S-Layer Proteins

2012 ◽  
Vol 195 (5) ◽  
pp. 977-989 ◽  
Author(s):  
J. M. Lunderberg ◽  
S.-M. Nguyen-Mau ◽  
G. S. Richter ◽  
Y.-T. Wang ◽  
J. Dworkin ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Secondary Cell Wall ◽  
Cell Wall Polysaccharide

scholarly journals Distinct Pathways Carry Out α and β Galactosylation of Secondary Cell Wall Polysaccharide in Bacillus anthracis

2020 ◽  
Vol 202 (15) ◽  
Author(s):  
Alice Chateau ◽  
So Young Oh ◽  
Anastasia Tomatsidou ◽  
Inka Brockhausen ◽  
Olaf Schneewind ◽  
...  
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Secondary Cell Wall ◽  
Turgor Pressure ◽  
Cell Wall Polysaccharide ◽  
Bacterial Cells ◽  
Content Type ◽  
Repeat Units ◽  
Undecaprenyl Phosphate

ABSTRACT Bacillus anthracis, the causative agent of anthrax disease, elaborates a secondary cell wall polysaccharide (SCWP) that is required for the retention of surface layer (S-layer) and S-layer homology (SLH) domain proteins. Genetic disruption of the SCWP biosynthetic pathway impairs growth and cell division. B. anthracis SCWP is comprised of trisaccharide repeats composed of one ManNAc and two GlcNAc residues with O-3–α-Gal and O-4–β-Gal substitutions. UDP-Gal, synthesized by GalE1, is the substrate of galactosyltransferases that modify the SCWP repeat. Here, we show that the gtsE gene, which encodes a predicted glycosyltransferase with a GT-A fold, is required for O-4–β-Gal modification of trisaccharide repeats. We identify a DXD motif critical for GtsE activity. Three distinct genes, gtsA, gtsB, and gtsC, are required for O-3–α-Gal modification of trisaccharide repeats. Based on the similarity with other three-component glycosyltransferase systems, we propose that GtsA transfers Gal from cytosolic UDP-Gal to undecaprenyl phosphate (C55-P), GtsB flips the C55-P-Gal intermediate to the trans side of the membrane, and GtsC transfers Gal onto trisaccharide repeats. The deletion of galE1 does not affect growth in vitro, suggesting that galactosyl modifications are dispensable for the function of SCWP. The deletion of gtsA, gtsB, or gtsC leads to a loss of viability, yet gtsA and gtsC can be deleted in strains lacking galE1 or gtsE. We propose that the loss of viability is caused by the accumulation of undecaprenol-bound precursors and present an updated model for SCWP assembly in B. anthracis to account for the galactosylation of repeat units. IMPORTANCE Peptidoglycan is a conserved extracellular macromolecule that protects bacterial cells from turgor pressure. Peptidoglycan of Gram-positive bacteria serves as a scaffold for the attachment of polymers that provide defined bacterial interactions with their environment. One such polymer, B. anthracis SCWP, is pyruvylated at its distal end to serve as a receptor for secreted proteins bearing the S-layer homology domain. Repeat units of SCWP carry three galactoses in B. anthracis. Glycosylation is a recurring theme in nature and often represents a means to mask or alter conserved molecular signatures from intruders such as bacteriophages. Several glycosyltransferase families have been described based on bioinformatics prediction, but few have been studied. Here, we describe the glycosyltransferases that mediate the galactosylation of B. anthracis SCWP.

scholarly journals Galactosylation of the Secondary Cell Wall Polysaccharide ofBacillus anthracisand Its Contribution to Anthrax Pathogenesis

2017 ◽  
Vol 200 (5) ◽  
Author(s):  
Alice Chateau ◽  
Justin Mark Lunderberg ◽  
So Young Oh ◽  
Teresa Abshire ◽  
Arthur Friedlander ◽  
...  
Keyword(s):  
Cell Wall ◽  
Glutamic Acid ◽  
Bacillus Anthracis ◽  
Secondary Cell Wall ◽  
Cell Wall Polysaccharide ◽  
Wild Type ◽  
Content Type ◽  
Bacterial Inoculum ◽  
Associated Proteins

ABSTRACTBacillus anthracis, the causative agent of anthrax disease, elaborates a secondary cell wall polysaccharide (SCWP) that is essential for bacterial growth and cell division.B. anthracisSCWP is comprised of trisaccharide repeats with the structure, [→4)-β-ManNAc-(1→4)-β-GlcNAc(O3-α-Gal)-(1→6)-α-GlcNAc(O3-α-Gal,O4-β-Gal)-(1→]6-12. The genes whose products promote the galactosylation ofB. anthracisSCWP are not yet known. We show here that the expression ofgalE1, encoding a UDP-glucose 4-epimerase necessary for the synthesis of UDP-galactose, is required forB. anthracisSCWP galactosylation. ThegalE1mutant assembles surface (S) layer and S layer-associated proteins that associate with ketal-pyruvylated SCWP via their S layer homology domains similarly to wild-typeB. anthracis, but the mutant displays a defect in γ-phage murein hydrolase binding to SCWP. Furthermore, deletion ofgalE1diminishes the capsulation ofB. anthraciswith poly-d-γ-glutamic acid (PDGA) and causes a reduction in bacterial virulence. These data suggest that SCWP galactosylation is required for the physiologic assembly of theB. anthraciscell wall envelope and for the pathogenesis of anthrax disease.IMPORTANCEUnlike virulentBacillus anthracisisolates,B. anthracisstrain CDC684 synthesizes secondary cell wall polysaccharide (SCWP) trisaccharide repeats without galactosyl modification, exhibits diminished growthin vitroin broth cultures, and is severely attenuated in an animal model of anthrax. To examine whether SCWP galactosylation is a requirement for anthrax disease, we generated variants ofB. anthracisstrains Sterne 34F2 and Ames lacking UDP-glucose 4-epimerase by mutating the genesgalE1andgalE2. We identifiedgalE1as necessary for SCWP galactosylation. Deletion ofgalE1decreased the poly-d-γ-glutamic acid (PDGA) capsulation of the vegetative form ofB. anthracisand increased the bacterial inoculum required to produce lethal disease in mice, indicating that SCWP galactosylation is indeed a determinant of anthrax disease.

α ε-Diaminopimelic Acid in the Peptide Moiety of the Cell Wall Polysaccharide of Bacillus anthracis

Nature ◽  
1956 ◽  
Vol 178 (4538) ◽  
pp. 865-866 ◽  
Author(s):  
H. SMITH ◽  
R. E. STRANGE ◽  
H. T. ZWARTOUW
Keyword(s):  
Cell Wall ◽  
Bacillus Anthracis ◽  
Diaminopimelic Acid ◽  
Cell Wall Polysaccharide ◽  
Peptide Moiety
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