Peptidoglycan synthesis by partly autolyzed cells of Bacillus subtilis W23

Harrington, Charles R.; Baddiley, J.

doi:10.1128/jb.155.2.776-792.1983

Cited by 12 publications

(12 citation statements)

References 59 publications

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“…The solvents and biochemicals used were of analytical grade and obtained from BDH Laboratories (Poole, UK). Tunicamycin was a gift from Dr G. Tamura and stored in alkaline solution as described [17]. Moraprenol-PP-GlcNAc was supplied by Dr V. N. Shibaev and Dr S. D. Maltsev as a methanolic solution containing ammonium acetate (30.7 ninol GlcNAc-I-phosphate ml-I).…”

Section: Materials and Analytical Proceduresmentioning

confidence: 99%

Biosynthesis of wall tiechoic acids in Staphylococcus aureus H, Micrococcus varians and Bacillus subtilis W23

Harrington

Baddiley

1985

European Journal of Biochemistry

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The precursors for linkage unit (LU) synthesis in Staphylococcus aureus H were UDP-GlcNAc, UDP-Nacetylmannosamine (ManNAc) and CDP-glycerol and synthesis was stimulated by ATP. Moraprenol-PPGlcNAc-ManNAc-(glycerol phosphate) -was formed from chemically synthesised nioraprenol-PP-GlcNAc, UDP-ManNAc and CDP-glycerol in the presence of Triton X-100. LU intermediates formed under both conditions served as acceptors for ribitol phosphate residues, from CDP-ribitol, which comprise the main chain. The initial transfer of GlcNAc-I-phosphate from UDP-GlcNAc was very sensitive to tunicamycin whereas the subsequent transfer of ManNAc from UDP-ManNAc was not.Poly(G1cNAc-I -phosphate) and LU synthesis in Micrococcus varians, with endogenous lipid acceptor, UDPGlcNAc and CDP-glycerol, was stimulated by UDP-ManNAc. Synthesis of LU on exogenous moraprenol-PPGlcNAc, with Triton X-100, was dependent on UDP-ManNAc and CDP-glycerol and the intermediates formed served as substrates for polymer synthesis. Membranes from Bacillus subtilis W23 had much lower levels of LU synthesis, but UDP-ManNAc was again required for optimal synthesis in the presence of UDP-GlcNAc and CDPglycerol. Conditions for LU synthesis on exogenous moraprenol-PP-GlcNAc were not found in this organism.LU synthesis on endogenous acceptor in the absence of UDP-ManNAc was explained by contamination of membranes with UDP-GlcNAc 2-epimerase. Under appropriate conditions, low levels of this enzyme were sufficient to convert UDP-GlcNAc into a mixture of UDP-Glc-NAc and UDP-ManNAc and account for LU Structural studies on the ribitol teichoic acids of Staphylococcus aureus H and Bacillus subtilis W23 and the poly-(GlcNAc-1-phosphate) of Micrococcus varians showed that these polymers are covalently attached to the muramic acid of peptidoglycan through a common linkage of (Gro-P),-GlcNAc-1-P [l]. The biosynthesis of teichoic acid and its linkage unit (LU) has been recently reviewed [2]. LU is synthesised from the precursors, UDP-GlcNAc and CDPglycerol, through a series of polyisoprenol pyrophosphate intermediates which have been isolated and characterised [3 -71. In S. uureus membranes, poly(ribito1 phosphate) may

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Section: Materials and Analytical Proceduresmentioning

confidence: 99%

Biosynthesis of wall tiechoic acids in Staphylococcus aureus H, Micrococcus varians and Bacillus subtilis W23

Harrington

Baddiley

1985

European Journal of Biochemistry

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“…ATP is synthesized inside the cell, whereas lipoteichoic acid seems to be concentrated in the outer layer of the cytoplasmic membrane (21 and references therein). Alanylation of lipoteichoic acid may therefore be a transmembrane process such as has previously been documented for the biosynthesis of teichoic acid (4) and peptidoglycan (13,14) in B. subtilis.…”

Section: Discussionmentioning

confidence: 77%

Maintenance of D-alanine ester substitution of lipoteichoic acid by reesterification in Staphylococcus aureus

Koch¹,

Doker²,

Fischer³

1985

J Bacteriol

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Toluene-treated Staphylococcus aureus cells did not synthesize teichoic acid and lipoteichoic acid under the conditions used. The organism displayed, however, a high capacity of incorporating D-[14C]alanine into previously formed polymers. The reaction was dependent on ATP and enhanced by magnesium ions. The incorporation rate into lipoteichoic acid correlated with the rate of loss of alanine ester which occurred through transfer to teichoic acid and base-catalyzed hydrolysis. At pH 6.5 the loss (20% within 4 h) was completely compensated for by reesterification. At pH 7.5 the loss was 60%, but by accelerated incorporation it was reduced to 10%. Incorporation was also enhanced when the original substitution of lipoteichoic acid was lowered by previous growth of S. aureus at high salt concentration. The newly added alanine was randomly distributed along the poly(glycerophosphate) chain. The decreased alanine substitution of lipoteichoic acid after growth at high salt concentration was shown to result from a direct inhibition of alanine incorporation. Lipoteichoic acid and teichoic acid are widespread membrane and cell wall components of gram-positive bacteria. In many cases both polymers are replaced with D-alanine ester (1, 3, 6). A concomitant decrease of the alanine content of teichoic acid (15) and lipoteichoic acid (10) in Staphylococcus aureus at increasing salt concentrations in the growth medium led us to hypothesize that the incorporation of D-alanine into both polymers may be under the same control. In Lactobacillus casei the incorporation into lipoteichoic acid was shown to occur by a two-step reaction (5, 19): Enzyme + D-alanine + ATP = enzyme alanylAMP + PPa Enzyme alanylAMP + lipoteichoic acid alanyl lipoteichoic acid + AMP + enzyme Recent pulse-chase experiments of growing S. aureus with D-[14C]alanine revealed a rapid turnover of the alanine ester

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“…We previously studied (22) the synthesis of peptidoglycan by intact cells undergoing partial wall autolysis and reached similar conclusions, although it was still possible that the wall-synthesizing complexes had been displaced from the membrane in the in vitro systems. Whereas displacement of the complexes does not occur in vivo, rotation of transmembrane complexes remains a plausible explanation for translocation in vivo; energy for such rotation or reorientation might arise from the electrocheniical proton gradient (AILH+) as previously suggested (22).…”

mentioning

confidence: 79%

“…In a preliminary report (23) we showed that peptidoglycan formation in B. subtilis W23 is inhibited by the uncouplers DNP and CCCP; this occurs with an accumulation of the nucleotide precursors involved in peptidoglycan synthesis. However, since these uncouplers also affect the transport of N-acetylglucosamine (GlcNAc) (23) and have been reported to initiate autolytic activity (26), the study was extended to the synthesis of teichoic acid in vivo. A glycerol auxotroph (34) was used to allow maximum incorporation of glycerol into teichoic acid.…”

mentioning

confidence: 90%

See 1 more Smart Citation

Synthesis of peptidoglycan and teichoic acid in Bacillus subtilis: role of the electrochemical proton gradient

Harrington¹,

Baddiley²

1984

J Bacteriol

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The effects of several ionophores and uncouplers on glycerol and N-acetylglucosamine incorporation by Bacillus subtilis 61360, a glycerol auxotroph, were tested at different pH values. In particular, the effect of valinomycin on the synthesis of teichoic acid and peptidoglycan was examined in pnore detail in both growing tells and in vitro biosynthetic systems. Valinomycin inhibited synthesis of wall teichoic acid and peptidoglycan in whole cells but not in the comparable in vitro systems. It did not inhiblt fornmation of free lipid or lipoteichoic acid. The results were consistent with a role for the electrochemical proton gradient in maintaining full activity of cell wall synthetic enzymes in intact cells. Such an energy source would be required for a niodel in which rotation or reorientation of synthetic enzyme complexes is envisaged for the translocation of wall precursor molecules across the cytoplasmic membrane (Harrington and Baddiley,

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Peptidoglycan synthesis by partly autolyzed cells of Bacillus subtilis W23

Cited by 12 publications

References 59 publications

Biosynthesis of wall tiechoic acids in Staphylococcus aureus H, Micrococcus varians and Bacillus subtilis W23

Biosynthesis of wall tiechoic acids in Staphylococcus aureus H, Micrococcus varians and Bacillus subtilis W23

Maintenance of D-alanine ester substitution of lipoteichoic acid by reesterification in Staphylococcus aureus

Synthesis of peptidoglycan and teichoic acid in Bacillus subtilis: role of the electrochemical proton gradient

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