Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Jun;130(3):1030–1037. doi: 10.1128/jb.130.3.1030-1037.1977

Initiation of Deoxyribonucleic Acid Replication in Germinating Spores of Bacillus subtilis 168 Carrying the dnaB(Ts)134 Mutation

Heather Callister 1, Sue Le Mesurier 1, R G Wake 1
PMCID: PMC235324  PMID: 405369

Abstract

The nature of the deoxyribonucleic acid synthesis reported by others to occur at 45°C in germinating spores of the temperature-sensitive deoxyribonucleic acid initiation mutant of Bacillus subtilis 168, TsB134, has been investigated. Density transfer experiments, using 5-bromouracil, show that a normal round of replication can occur in a significant fraction of the spore population under such conditions. No repair synthesis is detectable. The possibility raised by this finding, that initiation of the first round of replication during spore outgrowth is unique in that its initiation is determined prior to germination, has been investigated by comparing the behavior of germinating spores of isogenic strains of B. subtilis 168, one carrying and the other without the dnaB (Ts)134 mutation. It is shown that deoxyribonucleic acid synthesis in the Ts strain is very sensitive to temperature in the vicinity of 45°C. At a slightly higher temperature, 49°C, initiation of the first round of replication in the Ts strain is completely (greater than 96%) blocked, but it proceeds normally in the Ts+ strain. Thus, it is concluded that, after the germination of a spore, the action of the dnaB134 gene product is an obligatory requirement for initiation of the first round of replication. The initiation of replication that can occur in spores of the original TsB134 strain germinating at 45°C is presumably due to incomplete inactivation of the dnaB134 gene product under such conditions.

Full text

PDF
1030

Selected References

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

  1. Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burnett L., Wake R. G. Initiation and termination of chromosome replication at 45 degree C in a temperature-sensitive deoxyribonucleic acid initiation mutant of Bacillus subtilis 168, TsB134. J Bacteriol. 1977 Apr;130(1):538–539. doi: 10.1128/jb.130.1.538-539.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Callister H., Wake R. G. Completed chromosomes in thymine-requiring Bacillus subtilis spores. J Bacteriol. 1974 Nov;120(2):579–582. doi: 10.1128/jb.120.2.579-582.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Callister H., Wake R. G. Homogeneity in Bacillus subtilis spore DNA content. J Mol Biol. 1976 Apr 5;102(2):367–371. doi: 10.1016/s0022-2836(76)80060-5. [DOI] [PubMed] [Google Scholar]
  5. Hara H., Yoshikawa H. Asymmetric bidirectional replication of Bacillus subtilis chromosome. Nat New Biol. 1973 Aug 15;244(137):200–203. doi: 10.1038/newbio244200a0. [DOI] [PubMed] [Google Scholar]
  6. Karamata D., Gross J. D. Isolation and genetic analysis of temperature-sensitive mutants of B. subtilis defective in DNA synthesis. Mol Gen Genet. 1970;108(3):277–287. doi: 10.1007/BF00283358. [DOI] [PubMed] [Google Scholar]
  7. Laurent S. J., Vannier F. S. Temperature-sensitive initiation of chromosome replication in a mutant of Bacillus subtilis. J Bacteriol. 1973 May;114(2):474–484. doi: 10.1128/jb.114.2.474-484.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mastromei G., Riva S. Interconversion of thymine and thymidine in a thymine requiring strain of Bacillus subtilis. Biochem Biophys Res Commun. 1975 Mar 17;63(2):522–528. doi: 10.1016/0006-291x(75)90719-6. [DOI] [PubMed] [Google Scholar]
  9. Mendelson N. H. Can defective segregation prevent initiation? Cold Spring Harb Symp Quant Biol. 1968;33:313–316. doi: 10.1101/sqb.1968.033.01.035. [DOI] [PubMed] [Google Scholar]
  10. Mendelson N. H., Gross J. D. Characterization of a temperature-sensitive mutant of Bacillus subtilis defective in deoxyribonucleic acid replication. J Bacteriol. 1967 Nov;94(5):1603–1608. doi: 10.1128/jb.94.5.1603-1608.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Murakami S., Yoshikawa H. Gene that controls initiation of chromosome replication and prophage induction in Bacillus subtilis. Nature. 1976 Jan 22;259(5540):215–218. doi: 10.1038/259215a0. [DOI] [PubMed] [Google Scholar]
  12. Nukushina J. I., Ikeda Y. Genetic analysis of the developmental processes during germination and outgrowth of Bacillus subtilis spores with temperature-sensitive mutants. Genetics. 1969 Sep;63(1):63–74. doi: 10.1093/genetics/63.1.63. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. OISHI M., YOSHIKAWA H., SUEOKA N. SYNCHRONOUS AND DICHOTOMOUS REPLICATIONS OF THE BACILLUS SUBTILIS CHROMOSOME DURING SPORE GERMINATION. Nature. 1964 Dec 12;204:1069–1073. doi: 10.1038/2041069a0. [DOI] [PubMed] [Google Scholar]
  14. Upcroft P., Dyson H. J., Wake R. G. Characteristics of a Bacillus subtilis W23 mutant temperature sensitive for initiation of chromosome replication. J Bacteriol. 1975 Jan;121(1):121–127. doi: 10.1128/jb.121.1.121-127.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. WOESE C. R., FORRO J. R. Correlations between ribonucleic acid and deoxyribonucleic acid metabolism during spore germination. J Bacteriol. 1960 Dec;80:811–817. doi: 10.1128/jb.80.6.811-817.1960. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Wake R. G. A study of the possible extent of synthesis of repair DNA during germination of Bacillus subtilis spores. J Mol Biol. 1967 Apr 28;25(2):217–234. doi: 10.1016/0022-2836(67)90139-8. [DOI] [PubMed] [Google Scholar]
  17. White K., Sueoka N. Temperature-sensitive DNA synthesis mutants of Bacillus subtilis--appendix: theory of density transfer for symmetric chromosome replication. Genetics. 1973 Feb;73(2):185–214. doi: 10.1093/genetics/73.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yoshikawa H. Chromosomes in Bacillus subtilis spores and their segregation during germination. J Bacteriol. 1968 Jun;95(6):2282–2292. doi: 10.1128/jb.95.6.2282-2292.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES