1887

Abstract

A filamentous division mutant, PV302, of QM B1551 was isolated while screening for sporulation-defective mutants after nitrosoguanidine mutagenesis. Both phase-contrast and electron microscopy revealed that the mutant produced small spherical cells as well as filaments. It also accumulated large amounts of poly--hydroxybutyrate. Poly--hydroxybutyrate accounted for 16% of the dry weight of the mutant strain even after 28 h growth. In comparison to the parental strain, the division mutant also showed both an inability to sporulate and a reduced growth rate. All these phenotypes transduced together. Revertants gained the ability to sporulate, divide, and grow normally. Transductional mapping of the mutation, designated , established a new linkage group for consisting of and the pyrimidine biosynthesis genes . The spherical cells were separated from filaments by sucrose gradients and were tested for nucleic acid content and viability. The purified spherical cell fraction contained one-fifth the amount of DNA per mg protein as compared with the filamentous cell fraction and was shown to contain both non-viable minicells and some cells capable of growing after a lag of about 4 h. This suggests that the mutation not only causes defects in septum placement and sporulation, but may possibly affect DNA partitioning.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-136-3-545
1990-03-01
2021-09-24
Loading full text...

Full text loading...

/deliver/fulltext/micro/136/3/mic-136-3-545.html?itemId=/content/journal/micro/10.1099/00221287-136-3-545&mimeType=html&fmt=ahah

References

  1. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. 1967; Miniature Escherichia coli cells deficient in DNA. Proceedings of the National Academy of Sciences of the United States of America 57:321–326
    [Google Scholar]
  2. Bachmann B. 1983; Linkage map of Escherichia coli K12, edition 7. Microbiological Reviews 47:180–230
    [Google Scholar]
  3. Coyne S. I., Mendelson N. H. 1974; Clonal analysis of cell division in the Bacillus subtilis divIVB-1 minicell-producing mutant. Journal of Bacteriology 118:15–20
    [Google Scholar]
  4. Donachie W. D., Robinson A. C. 1987; Cell division: parameters values and process. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp 1578–1593 Neidhardt F. C., Ingraham J. L., Low K. B., Magasanik B., Schaecter M., Umbarger H. E. Edited by Washington, DC: American Society for Microbiology;
    [Google Scholar]
  5. Donachie W. D., Begg K. J., Sullivan N. F. 1984; Morphogenes of Escherichia coli. . In Microbial Development pp Lasick R., Shapiro L. Edited by Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  6. Donachie W. D., Kehoe M. 11984; The minicell system as a method for studying expression from plasmid DNA. Methods in Microbiology 17233–258
    [Google Scholar]
  7. English J. D., Vary P. S. 1986; Isolation of recombination-defective and UV-sensitive mutants of Bacillus megaterium. . Journal of Bacteriology 165:155–160
    [Google Scholar]
  8. Forsberg C. W., Costerton J. W., Macleod R. A. 1970; Separation and localization of cell wall layers of a gram-negative bacterium. Journal of Bacteriology 104:1338–1353
    [Google Scholar]
  9. Garbe J. C., Hess G. F., Franzen M. A., Vary P. S. 1984; Genetics of leucine biosynthesis in Bacillus megaterium QM B1551. Journal of Bacteriology 157:454–459
    [Google Scholar]
  10. Hirsh J. G., Fedorko M. F. 1968; Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and ‘post fixation’ in uranyl acetate. Journal of Cell Biology 38:615–627
    [Google Scholar]
  11. Hitchins A. D., Sadoff H. L. 1974; Properties of a thermosensitive asporogenous filamentous mutant of Bacillus megaterium. . Journal of Bacteriology 118:1167–1175
    [Google Scholar]
  12. Hitchins A. D., Slepecky R. A. 1969; Bacterial sporulation as a modified procaryotic cell division. Nature, London 223:804–807
    [Google Scholar]
  13. Janssen F. A., Lund A. J., Anderson L. E. 1958; Colorimetric assay for dipicolinic acid in bacterial spores. Science 127:26–27
    [Google Scholar]
  14. Kieselburg M. K., Weickert M., Vary P. S. 1984; Analysis of resident and transformant plasmids in Bacillus megaterium. . Biotechnology 2:254–259
    [Google Scholar]
  15. Law J. H., Slepecky R. A. 1961; Assay of poly-β-hydroxybutyr-ate. Journal of Bacteriology 82:33–36
    [Google Scholar]
  16. Love P. E., Yasbin R. E. 1986; Induction of the Bacillus subtilis SOS-like response by Escherichia coli RecA protein. Proceedings of the National Academy of Sciences of the United States of America 83:5204–5208
    [Google Scholar]
  17. Macrae R. M., Wilkinson J. F. 1958; Poly-β-hydroxybutyrate metabolism in washed suspensions of Bacillus cereus and Bacillus megaterium. . Journal of General Microbiology 19:210–222
    [Google Scholar]
  18. Mendelson N. H. 1982; Bacterial growth and division: genes, structures, forces, and clocks. Microbiological Reviews 46:341–375
    [Google Scholar]
  19. Miyakawa Y., Komano T. 1981; Study on the cell cycle of Bacillus subtilis using temperature-sensitive mutants. I. Isolation and genetic analysis of the mutants defective in septum formation. Molecular and General Genetics 181:207–214
    [Google Scholar]
  20. Mollenhauer H. H. 1963; Plastic embedding mixtures for use in electron microscopy. Stain Technology 39:111–114
    [Google Scholar]
  21. Mulder E., Woldringh C. L. 1989; Actively replicating nucleoids influence positioning of division sites in Escherichia coli filaments forming cells lacking DNA. Journal of Bacteriology 171:4303–4314
    [Google Scholar]
  22. Murray R.G.E., Gillen D. H., Heagy F. C. 1950; Cytological changes in Escherichia coli produced by infection with phage T2. Journal of Bacteriology 59:603–615
    [Google Scholar]
  23. Potvin B. W., Kelleher R. J., Gooder H. 1975; Pyrimidine biosynthetic pathway of Bacillus subtilis. . Journal of Bacteriology 123:604–615
    [Google Scholar]
  24. Reeve J. 1979; Use of minicells for bacteriophage-directed polypeptide synthesis. Methods in Enzymology 68:493–503
    [Google Scholar]
  25. Reeve J. N., Mendelson N. H., Coyne S. I., Hallock L. L., Cole R. M. 1973; Minicells of Bacillus subtilis. . Journal of Bacteriology 114:860–873
    [Google Scholar]
  26. Rotman Y., Fields M. L. 1968; A modified reagent for dipicolinic acid analysis. Analytical Biochemistry 22:168–172
    [Google Scholar]
  27. Sadie Y., Kada T. 1985; Bacillus subtilis gene involved in cell division, sporulation, and exoenzyme secretion. Journal of Bacteriology 163:648–653
    [Google Scholar]
  28. Schneider W. C. 1957; Determination of nucleic acids in tissues by pentose analysis. Methods in Enzymology 3:680–682
    [Google Scholar]
  29. Setlow J. K., Boling M. E., Allison D. P., Beattie K. L. 1973; The relationship between prophage induction and transformation in Haemophilis influenzae. . Journal of Bacteriology 115:153–161
    [Google Scholar]
  30. Shivakumar A. G., Hahn G. J., Dubnau D. 1979; Studies on the synthesis of plasmid-coded proteins and their control in Bacillus subtilis minicells. Plasmid 2:279–289
    [Google Scholar]
  31. Shivakumar A. G., Vanags R. I., Wilcox D. R., Katz L., Vary P. S., Fox J. L. 1989; Gene dosage effect on the expression of the delta-endotoxin genes of Bacillus thuringiensis subsp. kurstaki in Bacillus subtilis and Bacillus megaterium. . Gene 79:21–31
    [Google Scholar]
  32. Slepecky R. A., Law J. H. 1961; Synthesis and degradation of poly-β-hydroxybutyric acid in connection with sporulation of Bacillus megaterium. . Journal of Bacteriology 82:37–42
    [Google Scholar]
  33. Sussman M. D., Vary P. S., Hartman C., Setlow P. 1989; Integration and mapping of Bacillus megaterium genes which code for small, acid-soluble spore proteins and their protease. Journal of Bacteriology 170:4942–4945
    [Google Scholar]
  34. Vary P. S., Tao Y.-P. 1988; Development of genetic methods in Bacillus megaterium. . In Genetics and Biotechnology of Bacilli II pp 403–408 Hoch J. A., Ganesan A. T. Edited by San Diego: Academic Press;
    [Google Scholar]
  35. Van Alstyne D., Simon M. I. 1971; Division mutants of Bacillus subtilis: isolation and PBS1 transduction of division-specific markers. Journal of Bacteriology 108:1366–1379 of Bacillus megaterium. Journal of Bacteriology155
    [Google Scholar]
  36. Voros J., Goodman R. N. 1965; Filamentous forms of Erwinia amylovora. . Phytopathology 55:876–879
    [Google Scholar]
  37. Ward A. C., Dawes E. A. 1973; A disk assay for poly-β-hydroxybutyrate. Analytical Biochemistry 52:607–613
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-136-3-545
Loading
/content/journal/micro/10.1099/00221287-136-3-545
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error