Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1969 Feb;3(2):248–260. doi: 10.1128/jvi.3.2.248-260.1969

Defective Bacteriophage PBSH in Bacillus subtilis

II. Intracellular Development of the Induced Prophage

Martin Haas 1, Hiroshi Yoshikawa 1
PMCID: PMC375758  PMID: 4975369

Abstract

Treatment of Bacillus subtilis strain 168 with mitomycin C caused induction of a defective prophage, PBSH. During induction, extensive deoxyribonucleic acid (DNA) synthesis took place. Concurrently, a change in marker frequency of the bacterial DNA was noticed. The frequency of only one marker, ade-16, the marker closest to the origin of the bacterial chromosome, was enhanced manyfold. DNA from whole phage particles transformed all bacterial markers at a frequency equal to that of DNA in the noninduced culture, except ade-16, the frequency of which was enhanced 30 to 100 times. Analysis of a double isotope experiment demonstrated that 14% of the phage DNA was derived from preinduction bacterial DNA. The other 86% of DNA in phage particles was DNA replicated after induction. Density label experiments with 5-bromodeoxyuridine showed that postinduction DNA synthesis took place preferentially at the origin region of the bacterial chromosome. Measurement of the molecular weight of DNA replicated after induction clearly showed that postinduction DNA replication is chromosomal. No evidence for prophage detachment and autonomous phage DNA replication was found. The data indicated that, after mitomycin C action, the bacterial chromosome under-went multiple reinitiation at the origin, while normal sequential DNA replication was stopped. The pool of replicated bacterial DNA was fragmented randomly. This DNA was packaged into PBSH particles which were released after cell lysis.

Full text

PDF

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. BURGI E., HERSHEY A. D. Sedimentation rate as a measure of molecular weight of DNA. Biophys J. 1963 Jul;3:309–321. doi: 10.1016/s0006-3495(63)86823-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Ephrati-Elizur E. Spontaneous transformation in Bacillus subtilis. Genet Res. 1968 Feb;11(1):83–96. doi: 10.1017/s0016672300011216. [DOI] [PubMed] [Google Scholar]
  5. Frankel F. R. Evidence for long DNA strands in the replicating pool after T4 infection. Proc Natl Acad Sci U S A. 1968 Jan;59(1):131–138. doi: 10.1073/pnas.59.1.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Haas M., Yoshikawa H. Defective bacteriophage PBSH in Bacillus subtilis. I. Induction, purification, and physical properties of the bacteriophage and its deoxyribonucleic acid. J Virol. 1969 Feb;3(2):233–247. doi: 10.1128/jvi.3.2.233-247.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. IONESCO H., RYTER A., SCHAEFFER P. SUR UN BACT'ERIOPHAGE H'EBERG'E PAR LA SOUCHE MARBURG DE BACILLUS SUBTILIS. Ann Inst Pasteur (Paris) 1964 Dec;107:764–776. [PubMed] [Google Scholar]
  8. Iyer V. N. Mutations determining mitomycin resistance in Bacillus subtilis. J Bacteriol. 1966 Dec;92(6):1663–1669. doi: 10.1128/jb.92.6.1663-1669.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kammen H. O., Wojnar R. J., Canellakis E. S. Transformation in Bacillus subtilis. II. The development and maintenance of the competent state. Biochim Biophys Acta. 1966 Jul 20;123(1):56–65. [PubMed] [Google Scholar]
  10. Mahler I. Effect of mitomycin C on five excision-repair mutants of Bacillus subtilis. Biochem Biophys Res Commun. 1966 Oct 5;25(1):73–79. doi: 10.1016/0006-291x(66)90642-5. [DOI] [PubMed] [Google Scholar]
  11. Morse M L, Lederberg E M, Lederberg J. Transduction in Escherichia Coli K-12. Genetics. 1956 Jan;41(1):142–156. doi: 10.1093/genetics/41.1.142. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. O'Sullivan A., Sueoka N. Sequential replication of the Bacillus subtilis chromosome. IV. Genetic mapping by density transfer experiment. J Mol Biol. 1967 Jul 28;27(2):349–368. doi: 10.1016/0022-2836(67)90025-3. [DOI] [PubMed] [Google Scholar]
  13. PTASHNE M. THE DETACHMENT AND MATURATION OF CONSERVED LAMBDA PROPHAGE DNA. J Mol Biol. 1965 Jan;11:90–96. doi: 10.1016/s0022-2836(65)80174-7. [DOI] [PubMed] [Google Scholar]
  14. Skalka A., Butler B., Echols H. Genetic control of transcription during development of phage gamma. Proc Natl Acad Sci U S A. 1967 Aug;58(2):576–583. doi: 10.1073/pnas.58.2.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sueoka N., Yoshikawa H. The chromosome of Bacillus subtilis. I. Theory of marker frequency analysis. Genetics. 1965 Oct;52(4):747–757. doi: 10.1093/genetics/52.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. YOSHIKAWA H., SUEOKA N. Sequential replication of Bacillus subtilis chromosome. I. Comparison of marker frequencies in exponential and stationary growth phases. Proc Natl Acad Sci U S A. 1963 Apr;49:559–566. doi: 10.1073/pnas.49.4.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. YOSHIKAWA H., SUEOKA N. Sequential replication of the Bacillus subtilis chromosome. II. Isotopic transfer experiments. Proc Natl Acad Sci U S A. 1963 Jun;49:806–813. doi: 10.1073/pnas.49.6.806. [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]
  19. Yoshikawa H. DNA synthesis during germination of Bacillus subtilis spores. Proc Natl Acad Sci U S A. 1965 Jun;53(6):1476–1483. doi: 10.1073/pnas.53.6.1476. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yoshikawa H. Mutations Resulting from the Transformation of BACILLUS SUBTILIS. Genetics. 1966 Nov;54(5):1201–1214. doi: 10.1093/genetics/54.5.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yoshikawa H. The initiation of DNA replication in Bacillus subtilis. Proc Natl Acad Sci U S A. 1967 Jul;58(1):312–319. doi: 10.1073/pnas.58.1.312. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES