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. 1976 Jan;125(1):166–173. doi: 10.1128/jb.125.1.166-173.1976

Analysis of autolysins in temperature-sensitive morphological mutants of Bacillus subtilis.

W C Brown, C R Wilson, S Lukehart, F E Young, M A Shiflett
PMCID: PMC233348  PMID: 812860

Abstract

The content and distribution of autolysin were measured in temperature-sensitive morphological mutants of Bacillus subtilis. Strains RUB1000 and RUB1012 grew as rods at 30 C. At 45 C the mutants contained disproportionately less teichoic acid than peptidoglycan and grew as irregular spheres. The amount of enzyme that could be extracted from rods was at least 31 times the amount extracted from spheres. The rate of autolysis of cell walls was 7- to 28-fold greater in rods than in spheres. The low activity found associated with the cell walls of spheres was not compensated for by larger amounts of autolytic activity in the cytoplasm. No activity was found in the growth medium at either temperature. The failure of the mutant cells to autolyze was due to low amidase activity and relatively resistant cell walls. Revertants of RUB1012 were isolated that had 13, 23, and 55% of the normal proportions of teichoic acid when grown at the nonpermissive temperature. Cell walls from the revertants were as sensitive to added amidase as the wild-type strain. None of the revertant strains regained the wild-type ability to produce more amidase at 45 C. However, the deficiency in autolysin observed with RUB1012 was partially restored in revertants containing higher proportions of teichoic acid.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Boylan R. J., Mendelson N. H., Brooks D., Young F. E. Regulation of the bacterial cell wall: analysis of a mutant of Bacillus subtilis defective in biosynthesis of teichoic acid. J Bacteriol. 1972 Apr;110(1):281–290. doi: 10.1128/jb.110.1.281-290.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boylan R. J., Mendelson N. H. Initial characterization of a temperature-sensitive rod--mutant of Bacillus subtilis. J Bacteriol. 1969 Dec;100(3):1316–1321. doi: 10.1128/jb.100.3.1316-1321.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown W. C., Fraser D. K., Young F. E. Problems in purification of a Bacillus subtilis autolytic enzyme caused by association with teichoic acid. Biochim Biophys Acta. 1970 Feb 11;198(2):308–315. doi: 10.1016/0005-2744(70)90063-x. [DOI] [PubMed] [Google Scholar]
  4. Brown W. C. Rapid methods for extracting autolysins from Bacillus subtilis. Appl Microbiol. 1973 Feb;25(2):295–300. doi: 10.1128/am.25.2.295-300.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown W. C., Young F. E. Dynamic interactions between cell wall polymers, extracellular proteases and autolytic enzymes. Biochem Biophys Res Commun. 1970 Feb 20;38(4):564–568. doi: 10.1016/0006-291x(70)90618-2. [DOI] [PubMed] [Google Scholar]
  6. Chatterjee A. N., Mirelman D., Singer H. J., Park J. T. Properties of a novel pleiotropic bacteriophage-resistant mutant of Staphylococcus aureus H. J Bacteriol. 1969 Nov;100(2):846–853. doi: 10.1128/jb.100.2.846-853.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cole R. M., Popkin T. J., Boylan R. J., Mendelson N. H. Ultrastructure of a temperature-sensitive rod- mutant of Bacillus subtilis. J Bacteriol. 1970 Sep;103(3):793–810. doi: 10.1128/jb.103.3.793-810.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davie J. M., Brock T. D. Effect of teichoic acid on resistance to the membrane-lytic agent of Streptococcus zymogenes. J Bacteriol. 1966 Dec;92(6):1623–1631. doi: 10.1128/jb.92.6.1623-1631.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dubnau D., Goldthwaite C., Smith I., Marmur J. Genetic mapping in Bacillus subtilis. J Mol Biol. 1967 Jul 14;27(1):163–185. doi: 10.1016/0022-2836(67)90358-0. [DOI] [PubMed] [Google Scholar]
  10. Fan D. P., Beckman M. M., Cunningham W. P. Ultrastructural studies on a mutant of Bacillus subtilis whose growth is inhibited due to insufficient autolysin production. J Bacteriol. 1972 Mar;109(3):1247–1257. doi: 10.1128/jb.109.3.1247-1257.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fan D. P., Beckman M. M. Mutant of Bacillus subtilis demonstrating the requirement of lysis for growth. J Bacteriol. 1971 Feb;105(2):629–636. doi: 10.1128/jb.105.2.629-636.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fan D. P., Beckmann M. M. Mutant of Bacillus subtilis with a temperature-sensitive autolytic amidase. J Bacteriol. 1973 May;114(2):798–803. doi: 10.1128/jb.114.2.798-803.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Forsberg C. W., Rogers H. J. Characterization of Bacillus licheniformis 6346 mutants which have altered lytic enzyme activities. J Bacteriol. 1974 May;118(2):358–368. doi: 10.1128/jb.118.2.358-368.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Forsberg C. W., Wyrick P. B., Ward J. B., Rogers H. J. Effect of phosphate limitation on the morphology and wall composition of Bacillus licheniformis and its phosphoglucomutase-deficient mutants. J Bacteriol. 1973 Feb;113(2):969–984. doi: 10.1128/jb.113.2.969-984.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Forsberg C., Rogers H. J. Autolytic enzymes in growth of bacteria. Nature. 1971 Jan 22;229(5282):272–273. doi: 10.1038/229272a0. [DOI] [PubMed] [Google Scholar]
  16. Glaser L. Bacterial cell surface polysaccharides. Annu Rev Biochem. 1973;42:91–112. doi: 10.1146/annurev.bi.42.070173.000515. [DOI] [PubMed] [Google Scholar]
  17. Herbold D. R., Glaser L. Bacillus subtilis N-acetylmuramic acid L-alanine amidase. J Biol Chem. 1975 Mar 10;250(5):1676–1682. [PubMed] [Google Scholar]
  18. Higgins M. L., Shockman G. D. Procaryotic cell division with respect to wall and membranes. CRC Crit Rev Microbiol. 1971 May;1(1):29–72. doi: 10.3109/10408417109104477. [DOI] [PubMed] [Google Scholar]
  19. Krulwich T. A., Ensign J. C. Activity of an autolytic N-acetylmuramidase during sphere-rod morphogenesis in Arthrobacter crystallopoietes. J Bacteriol. 1968 Sep;96(3):857–859. doi: 10.1128/jb.96.3.857-859.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Nester E W, Schafer M, Lederberg J. Gene Linkage in DNA Transfer: A Cluster of Genes Concerned with Aromatic Biosynthesis in Bacillus Subtilis. Genetics. 1963 Apr;48(4):529–551. doi: 10.1093/genetics/48.4.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pooley H. M., Shockman G. D., Higgins M. L., Porres-Juan J. Some properties of two autolytic-defective mutants of Streptococcus faecalis ATCC 9790. J Bacteriol. 1972 Jan;109(1):423–431. doi: 10.1128/jb.109.1.423-431.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rogers H. J., McConnell M., Burdett I. D. Cell wall or membrane mutants of Bacillus subtilis and Bacillus licheniformis with grossly deformed morphology. Nature. 1968 Jul 20;219(5151):285–288. doi: 10.1038/219285a0. [DOI] [PubMed] [Google Scholar]
  24. Rogers H. J., McConnell M., Burdett I. D. The isolation and characterization of mutants of Bacillus subtilis and Bacillus licheniformis with disturbed morphology and cell division. J Gen Microbiol. 1970 May;61(2):155–171. doi: 10.1099/00221287-61-2-155. [DOI] [PubMed] [Google Scholar]
  25. Rogers H. J., McConnell M., Hughes R. C. The chemistry of the cell walls of rod mutants of Bacillus subtilis. J Gen Microbiol. 1971 Jun;66(3):297–308. doi: 10.1099/00221287-66-3-297. [DOI] [PubMed] [Google Scholar]
  26. Tomasz A. Biological consequences of the replacement of choline by ethanolamine in the cell wall of Pneumococcus: chanin formation, loss of transformability, and loss of autolysis. Proc Natl Acad Sci U S A. 1968 Jan;59(1):86–93. doi: 10.1073/pnas.59.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tomasz A. Choline in the cell wall of a bacterium: novel type of polymer-linked choline in Pneumococcus. Science. 1967 Aug 11;157(3789):694–697. doi: 10.1126/science.157.3789.694. [DOI] [PubMed] [Google Scholar]
  28. YOUNG F. E., SPIZIZEN J., CRAWFORD I. P. BIOCHEMICAL ASPECTS OF COMPETENCE IN THE BACILLUS SUBTILIS TRANSFORMATION SYSTEM. I. CHEMICAL COMPOSITION OF CELL WALLS. J Biol Chem. 1963 Sep;238:3119–3125. [PubMed] [Google Scholar]
  29. Young F. E. Autolytic enzyme associated with cell walls of Bacillus subtilis. J Biol Chem. 1966 Aug 10;241(15):3462–3467. [PubMed] [Google Scholar]
  30. Young F. E. Fractionation and partial characterization of the products of autolysis of cell walls of Bacillus subtilis. J Bacteriol. 1966 Oct;92(4):839–846. doi: 10.1128/jb.92.4.839-846.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]

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