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Volume 146, Issue 4

Incorporation of [2-H]glycerol into cell surface components of 168 and thermosensitive mutants affected in wall teichoic acid synthesis: effect of tunicamycin Free

Abstract

A method is described for measuring the synthesis of poly(glycerol phosphate) [poly(groP)], the major wall teichoic acid (WTA), lipoteichoic acid (LTA) and phospholipid (P-lipid), through fractionation of [2-H]glycerol ([2-H]gro)-labelled cells. When cultures of certain temperature-sensitive mutants defective in one of several genes, encoding enzymes involved in WTA synthesis, were transferred to the restrictive temperature, the synthesis of WTA underwent a specific, immediate, block, while that of LTA or P-lipid proceeded unimpeded. These results, in addition to confirming the role of genes, demonstrated, reciprocally, the specificity of the fractionation procedure used to distinguish label in WTA from that in LTA or P-lipid. Results of analysis of other, less severely affected, -deficient mutants, as well as of another genetically unrelated mutant developing comparable morphological phenotypes in non-permissive conditions, are discussed in relation to a possible mechanism generating the latter phenotype. Fractionation of 168 cells labelled either with [2-H]gro or with [1-C]-acetylglucosamine, to which tunicamycin was added at 05 μg ml (the MIC) revealed a specific and marked inhibition of poly(groP) as well as of poly(3--β-D-glucopyranosyl--acetylgalactosamine 1-phosphate), the minor WTA. However, for 60 min at least, the syntheses of PG, LTA and P-lipid were barely affected.

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Keyword(s): [2-3H]gro, [2-3H]glycerol , GlcNAc, N-acetylglucosamine , LTA, lipoteichoic acid , ND , nephelometric density , P-lipid, phospholipid , PG, peptidoglycan , phospholipid , poly(GlcGalNAc 1-P), poly(3-O-β-D-glucopyranosyl-N-acetylgalactosamine 1-phosphate) , poly(glycerol phosphate) , poly(groP), poly(glycerol phosphate) , teichoic acid , thermosensitive tag mutants , Ts, temperature sensitive , TUN, tunicamycin , tunicamycin and WTA, wall teichoic acid
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2000-04-01
2024-04-23
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Incorporation of [2-3H]glycerol into cell surface components of Bacillus subtilis 168 and thermosensitive mutants affected in wall teichoic acid synthesis: effect of tunicamycin
Microbiology 146, 797 (2000); https://doi.org/10.1099/00221287-146-4-797
/content/journal/micro/10.1099/00221287-146-4-797
/content/journal/micro/10.1099/00221287-146-4-797
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