1887

Abstract

The upstream region of the -acetylmuramoyl-L-alanine amidase gene (; a major autolysin) was cloned into by chromosome walking. Sequencing of the region showed the presence of two open reading frames, one (designated as ) which starts at a UUG codon and encodes a polypeptide of 705 amino acids with an of 76725, and the other (designated as ), upstream of , comprising 102 amino acids and having a signal sequence characteristic of a lipoprotein. Purification of the CwbA protein and determination of its N-terminal amino acid sequence revealed that it contains a presumed signal peptide which is processed after Ala at position 25 from the N-terminal, and that the of the mature form is 75000. The amino acid sequences of the N-terminal and C-terminal regions of CwbA were found to be highly homologous with those of the cell wall binding domain of CwlB and the gene product, respectively. CwbA stimulated the major autolysin activity approximately threefold . These data indicate that CwbA is the modifier protein of the major autolysin reported by Herbold, D. R. & Glaser, L. (1975; 250, 1676–1682). In-frame fusion between the and genes demonstrated that is translated and expressed during the exponential growth phase.

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1992-06-01
2021-05-10
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References

  1. Akamatsu T., Sekiguchi J. 1987; Genetic mapping and properties of filamentous mutations in Bacillus subtilis . Agricultural and Biological Chemistry 51:2901–2909
    [Google Scholar]
  2. Coûte J. G. 1972; Sporulation in Bacillus subtilis. Characterization of oligosporogenous mutants and comparison of their phenotypes with those of asporogenous mutants. Journal of General Microbiology 71:1–15
    [Google Scholar]
  3. Foster S. J. 1991; Cloning, expression, sequence analysis and biochemical characterization of autolytic amidase of Bacillus subtilis 168 trpC2 . Journal of General Microbiology 137:1987–1998
    [Google Scholar]
  4. Guinand M., Michel G., Balassa G. 1976; Lytic enzymes in sporulating Bacillus subtilis . Biochemical and Biophysical Research Communications 68:1287–1293
    [Google Scholar]
  5. Herbold D. R., Glaser L. 1975; Bacillus subtilis Af-acetylmuramic acid L-alanine amidase. Journal of Biological Chemistry 250:1676–1682
    [Google Scholar]
  6. Illing N., Errington J. 1991; Genetic regulation of morphogenesis in Bacillus subtilis: roles of σN and <rF in prespore engulfment. Journal of Bacteriology 173:3159–3169
    [Google Scholar]
  7. Kuroda A., Sekiguchi J. 1990; Cloning, sequencing and genetic mapping of a Bacillus subtilis cell wall hydrolase gene. Journal of General Microbiology 136:2209–2216
    [Google Scholar]
  8. Kuroda A., Sekiguchi J. 1991; Molecular cloning and sequencing of a major Bacillus subtilis autolysin gene. Journal of Bacteriology 173:7304–7312
    [Google Scholar]
  9. Lopez-Diaz I., Clarke S., Mandelstam J. 1986; spoIID operon of Bacillus subtilis: cloning and sequence. Journal of General Microbiology 132:341–354
    [Google Scholar]
  10. Mandel M., Higa A. 1970; Calcium-dependent bacteriophage DNA infection. Journal of Molecular Biology 53:159–162
    [Google Scholar]
  11. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning, a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  12. Miller J. H. 1972 Experiments in Molecular Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  13. Murao S., Takahara Y. 1974; Enzymes lytic against Pseudomonas aeruginosa produced by Bacillus subtilis YT-25. Agricultural and Biological Chemistry 38:2305–2316
    [Google Scholar]
  14. Okada S., Kitahata S. 1973; Purification and some properties of bacterial lysozyme. Journal of Fermentation Technology 51:705–712
    [Google Scholar]
  15. Rogers H. J., Taylor C, Rayter S., Ward J. B. 1984; Purification and properties of autolytic endo-β-N-acetyl-glucos-aminidase and the N-acetylmuramyl-L-alanine amidase from Bacillus subtilis strain 168. Journal of General Microbiology 130:2395–2402
    [Google Scholar]
  16. Sekiguchi J., Ezaki B., Kodama K., Akamatsu T. 1988; Molecular cloning of a gene affecting the autolysin level and flagellation in Bacillus subtilis . Journal of General Microbiology 134:1611–1621
    [Google Scholar]
  17. Shimotsu H., Henner D. J. 1986; Modulation of Bacillus subtilis levansucrase gene expression by sucrose and regulation of the steady-state mRNA level by sacU and sacQ genes. Journal of Bacteriology 168:380–388
    [Google Scholar]
  18. Stragier P. 1989; Temporal and spatial control of gene expression during sporulation: from facts to speculations. Regulation of Procaryotic Development243–254 Smith I., Slepecky R. A., Setlow P. Washington DC: American Society for Microbiology;
    [Google Scholar]
  19. Turner S. M., Mandelstam J. 1986; Cloning and sequencing of a gene from Bacillus amyloliquefaciens that complements mutations of the sporulation gene spoIID in Bacillus subtilis . Journal of General Microbiology 132:3025–3035
    [Google Scholar]
  20. Wu H. C., Tokunaga M. 1986; Biogenesis of lipoproteins in bacteria. Current Topics in Microbiology and Immunology 125:127–157
    [Google Scholar]
  21. Yanisch-Perron C, Vieira J., Messing J. 1985; Improved M13 phage cloning vectors and host strains : nucleotide sequences of the M13 mpl8 and pUC19 vectors. Gene 33:103–119
    [Google Scholar]
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