1887

Abstract

The operon of 168 encodes enzymes responsible for the synthesis of poly(glucosyl -acetylgalactosamine 1-phosphate) [poly(GlcGalNAc 1-P)], a wall teichoic acid (WTA). Analysis of the nucleotide sequence revealed that both GgaA and GgaB contained the motif characteristic of sugar transferases, while GgaB was most likely to be bifunctional, being endowed with an additional motif present in glucosyl/glycerophosphate transferases. Transcription of the operon was thermosensitive, and took place from an unusually distant -controlled promoter. The incorporation of the poly(GlcGalNAc 1-P) precursors by various mutants deficient in the synthesis of poly(glycerol phosphate), which is the most abundant WTA of strain 168, revealed that both WTAs were most likely to be attached to peptidoglycan (PG) through the same linkage unit (LU). The incorporation of poly(GlcGalNAc 1-P) precursors by protoplasts confirmed the existence of this LU, and provided further evidence that incorporation takes place at the outer surface of the protoplast membrane. The data presented here strengthen the view that biosynthesis of the LU, and the hooking of the LU-endowed polymer to PG, offer distinct widespread targets for antibiotics specific to Gram-positive bacteria.

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2006-06-01
2019-10-18
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References

  1. Antoniewski, C., Savelli, B. & Stragier, P. ( 1990; ). The spoIIJ gene, which regulates early developmental steps in Bacillus subtilis, belongs to a class of environmentally responsive genes. J Bacteriol 172, 86–93.
    [Google Scholar]
  2. Araki, Y. & Ito, E. ( 1989; ). Linkage units in cell walls of Gram-positive bacteria. Crit Rev Microbiol 17, 121–135.[CrossRef]
    [Google Scholar]
  3. Bertram, K. C., Hancock, I. C. & Baddiley, J. ( 1981; ). Synthesis of teichoic acid by Bacillus subtilis protoplasts. J Bacteriol 148, 406–412.
    [Google Scholar]
  4. Bhavsar, A. P., Erdman, L. K., Schertzer J. W. & Brown, E. D. ( 2004; ). Teichoic acid is an essential polymer in Bacillus subtilis that is functionally distinct from teichuronic acid. J Bacteriol 186, 7865–7873.[CrossRef]
    [Google Scholar]
  5. Chung, C. T. & Miller, R. H. ( 1988; ). A rapid and convenient method for the preparation and storage of competent bacterial cells. Nucleic Acids Res 16, 3580.[CrossRef]
    [Google Scholar]
  6. Del Sal, G., Manfioletti, G. & Schneider, C. ( 1988; ). A one-tube plasmid DNA mini-preparation suitable for sequencing. Nucleic Acids Res 16, 9878.[CrossRef]
    [Google Scholar]
  7. Estrela, A. I., Pooley, H. M., de Lencastre, H. & Karamata, D. ( 1991; ). Genetic and biochemical characterization of Bacillus subtilis 168 mutants specifically blocked in the synthesis of the teichoic acid poly(3-O-β-d-glucopyranosyl-N-acetylgalactosamine 1-phosphate): gneA, a new locus, is associated with UDP-N-acetylglucosamine 4-epimerase activity. J Gen Microbiol 137, 943–950.[CrossRef]
    [Google Scholar]
  8. Fitzgerald, S. N. & Foster, T. J. ( 2000; ). Molecular analysis of the tagF gene, encoding CDP-glycerol : poly(glycerophosphate) glycerophosphotransferase of Staphylococcus epidermidis ATCC 14990. J Bacteriol 182, 1046–1052.[CrossRef]
    [Google Scholar]
  9. Hanahan, D. ( 1983; ). Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166, 557–580.[CrossRef]
    [Google Scholar]
  10. Harrington, C. R. & Baddiley, J. ( 1985; ). Biosynthesis of wall teichoic acids in Staphylococcus aureus H, Micrococcus varians and Bacillus subtilis W23. Involvement of lipid intermediates containing the disaccharide N-acetylmannosaminyl N-acetylglucosamine. Eur J Biochem 153, 639–645.[CrossRef]
    [Google Scholar]
  11. Helmann, J. D. & Moran, C. P. ( 2002; ). RNA polymerase and sigma factors. In Bacillus subtilis and its Closest Relatives: from Genes to Cells, pp. 289–312. Edited by A. L. Sonenshein, J. A. Hoch & R. Losick. Washington, DC: American Society for Microbiology.
  12. Karamata, D. & Gross, J. D. ( 1970; ). Isolation and genetic analysis of temperature-sensitive mutants of B. subtilis defective in DNA synthesis. Mol Gen Genet 108, 277–287.
    [Google Scholar]
  13. Karamata, D., Pooley, H. M. & Monod, M. ( 1987; ). Expression of heterologous genes for wall teichoic acid in Bacillus subtilis 168. Mol Gen Genet 207, 73–81.[CrossRef]
    [Google Scholar]
  14. Kunst, F., Ogasawara, N., Moszer, I. & 148 other authors ( 1997; ). The complete genome sequence of the Gram-positive bacterium Bacillus subtilis. Nature 390, 249–256.[CrossRef]
    [Google Scholar]
  15. Lazarevic, V. & Karamata, D. ( 1995; ). The tagGH operon of Bacillus subtilis 168 encodes a two-component ABC transporter involved in the metabolism of two wall teichoic acids. Mol Microbiol 16, 345–355.[CrossRef]
    [Google Scholar]
  16. Lazarevic, V., Margot, P., Soldo, B. & Karamata, D. ( 1992; ). Sequencing and analysis of the Bacillus subtilis lytRABC divergon: a regulatory unit encompassing the structural genes of the N-acetylmuramoyl-l-alanine amidase and its modifier. J Gen Microbiol 138, 1949–1961.[CrossRef]
    [Google Scholar]
  17. Lazarevic, V., Mauël, C., Soldo, B., Freymond, P. P., Margot, P. & Karamata, D. ( 1995; ). Sequence analysis of the 308 degrees to 311 degrees segment of the Bacillus subtilis 168 chromosome, a region devoted to cell wall metabolism, containing non-coding grey holes which reveal chromosomal rearrangements. Microbiology 141, 329–335.[CrossRef]
    [Google Scholar]
  18. Lazarevic, V., Abellan, F.-X., Beggah-Möller, S., Karamata, D. & Mauël, C. ( 2002a; ). Comparison of ribitol and glycerol teichoic acid genes in Bacillus subtilis W23 and 168: identical function, similar divergent organization, but different regulation. Microbiology 148, 815–824.
    [Google Scholar]
  19. Lazarevic, V., Pooley, H. M., Mauël, C. & Karamata, D. ( 2002b; ). Teichoic and teichuronic acids from Gram-positive bacteria. In Biopolymers, vol. 5, Polysaccharides I: Polysaccharides from Prokaryotes, pp. 465–492. Edited by E. J. Vandamme, S. de Baets & A. Steinbüchel. Weinheim: Wiley-VCH.
  20. Longchamp, P. F., Mauël, C. & Karamata, D. ( 1994; ). Lytic enzymes associated with defective prophages of Bacillus subtilis: sequencing and characterization of the region comprising the N-acetylmuramoyl-l-alanine amidase gene of prophage PBSX. Microbiology 140, 1855–1867.[CrossRef]
    [Google Scholar]
  21. Marchler-Bauer, A., Anderson, J. B., DeWeese-Scott, C. & 24 other authors ( 2003; ). CDD: a curated Entrez database of conserved domain alignments. Nucleic Acids Res 31, 383–387.[CrossRef]
    [Google Scholar]
  22. Mauël, C., Young, M. & Karamata, D. ( 1991; ). Genes concerned with synthesis of poly(glycerol phosphate), the essential teichoic acid in Bacillus subtilis strain 168, are organized in two divergent transcription units. J Gen Microbiol 137, 929–941.[CrossRef]
    [Google Scholar]
  23. Mauël, C., Young, M., Monsutti-Grecescu, A., Marriott, S. A. & Karamata, D. ( 1994; ). Analysis of Bacillus subtilis tag gene expression using transcriptional fusions. Microbiology 140, 2279–2288.[CrossRef]
    [Google Scholar]
  24. Oultram, J. D., Peck, H., Brehm, J. K., Thompson, D. E., Swinfield, T. J. & Minton, N. P. ( 1988; ). Introduction of genes for leucine biosynthesis from Clostridium pasteurianum into C. acetobutylicum by cointegrate conjugal transfer. Mol Gen Genet 214, 177–179.[CrossRef]
    [Google Scholar]
  25. Pooley, H. M. & Karamata, D. ( 1988; ). Can synthesis of cell wall anionic polymers in Bacillus subtilis be a target for antibiotics? In Antibiotic Inhibition of Bacterial Cell Surface Assembly and Function, pp. 591–594. Edited by P. Actor, L. Daneo-Moore, M. L. Higgins, M. R. J. Salton & G. D. Shockman. Washington, DC: American Society for Microbiology.
  26. Pooley, H. M. & Karamata, D. ( 2000; ). 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, 797–805.
    [Google Scholar]
  27. Pooley, H. M., Paschoud, D. & Karamata, D. ( 1987; ). The gtaB marker in Bacillus subtilis 168 is associated with a deficiency in UDPglucose pyrophosphorylase. J Gen Microbiol 133, 3481–3493.
    [Google Scholar]
  28. Pooley, H. M., Abellan, F.-X. & Karamata, D. ( 1991; ). A conditional-lethal mutant of Bacillus subtilis 168 with a thermosensitive glycerol-3-phosphate cytidylyltransferase, an enzyme specific for the synthesis of the major wall teichoic acid. J Gen Microbiol 137, 921–928.[CrossRef]
    [Google Scholar]
  29. Pooley, H. M., Abellan, F.-X. & Karamata, D. ( 1992; ). CDP-glycerol : poly(glycerophosphate) glycerophosphotransferase, which is involved in the synthesis of the major wall teichoic acid in Bacillus subtilis 168, is encoded by tagF (rodC). J Bacteriol 174, 646–649.
    [Google Scholar]
  30. Rosenberger, R. F. ( 1976; ). Control of teichoic and teichuronic acid biosynthesis in Bacillus subtilis 168trp. Evidence for repression of enzyme synthesis and inhibition of enzyme activity. Biochim Biophys Acta 428, 516–524.[CrossRef]
    [Google Scholar]
  31. Shibaev, V. N., Duckworth, M., Archibald, A. R. & Baddiley, J. ( 1973; ). The structure of a polymer containing galactosamine from walls of Bacillus subtilis 168. Biochem J 135, 383–384.
    [Google Scholar]
  32. Soldo, B., Lazarevic, V., Margot, P. & Karamata, D. ( 1993; ). Sequencing and analysis of the divergon comprising gtaB, the structural gene of UDP-glucose pyrophosphorylase of Bacillus subtilis 168. J Gen Microbiol 139, 3185–3195.[CrossRef]
    [Google Scholar]
  33. Soldo, B., Lazarevic, V., Pagni, M. & Karamata, D. ( 1999; ). Teichuronic acid operon of Bacillus subtilis 168. Mol Microbiol 31, 795–805.[CrossRef]
    [Google Scholar]
  34. Soldo, B., Lazarevic, V. & Karamata, D. ( 2002; ). tagO is involved in the synthesis of all anionic cell-wall polymers in Bacillus subtilis 168. Microbiology 148, 2079–2087.
    [Google Scholar]
  35. Soldo, B., Scotti, C., Karamata, D. & Lazarevic, V. ( 2003; ). The Bacillus subtilis Gne (GneA, GalE) protein can catalyse UDP-glucose as well as UDP-N-acetylglucosamine 4-epimerisation. Gene 319, 65–69.[CrossRef]
    [Google Scholar]
  36. Yanisch-Perron, C., Vieira, J. & Messing, J. ( 1985; ). Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33, 103–119.[CrossRef]
    [Google Scholar]
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