Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1974 Dec;14(6):1369–1379. doi: 10.1128/jvi.14.6.1369-1379.1974

Inhibition of Bacteriophage φX174 DNA Replication in dnaB Mutants of Escherichia coli C

Lawrence B Dumas 1, Christine A Miller 1
PMCID: PMC355664  PMID: 4610183

Abstract

Bacteriophage φX174 DNA replication was examined in temperature-sensitive dnaB mutants of Escherichia coli C to determine which stages require the participation of the product of this host gene. The conversion of the infecting phage single-stranded DNA to the double-stranded replicative form (parental RF synthesis) is completely inhibited at the nonpermissive temperature (41 C) in two of the three dnaB mutants tested. The efficiency of phage eclipse and of phage DNA penetration of these mutant host cells at 41 C is the same as that of the parent host strain. The defect is most likely in the synthesis of the complementary strand DNA. The semiconservative replication of the double-stranded replicative form DNA (RF replication) is inhibited in all three host mutants after shifting from 30 to 41 C. Late in infection, the rate of progeny single-stranded phage DNA synthesis increases following shifts from 30 to 41 C. Approximately the same amounts of phage DNA and of infectious phage particles are made following the shift to 41 C as in the control left at 30 C. The simplest interpretation of our data is that the product of the host dnaB gene is required for φX174 parental RF synthesis and RF replication, but is not directly involved in phage single-stranded DNA synthesis once it has begun. The possible significance of the synthesis of parental RF DNA at 41 C in one of the three mutants is discussed.

Full text

PDF
1369

Selected References

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

  1. Bone D. R., Dowell C. E. A mutant of bacteriophage øX174 which infects E. coli K12 strains. II. Replication of øXtB in tsDNA strains. Virology. 1973 Apr;52(2):330–336. doi: 10.1016/0042-6822(73)90327-9. [DOI] [PubMed] [Google Scholar]
  2. Denhardt D. T., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. 3. Phage maturation and lysis after synchronized infection. J Mol Biol. 1965 Jul;12(3):641–646. doi: 10.1016/s0022-2836(65)80318-7. [DOI] [PubMed] [Google Scholar]
  3. Dumas L. B., Darby G., Sinsheimer R. L. The replication of bacteriophage phi X174 DNA in vitro. Temperature effects on repair synthesis and displacement synthesis. Biochim Biophys Acta. 1971 Jan 28;228(2):407–422. [PubMed] [Google Scholar]
  4. Dumas L. B., Miller C. A. Replication of bacteriophage phiX174 DNA in a temperature-sensitive dnaE mutant of Escherichia coli C. J Virol. 1973 Jun;11(6):848–855. doi: 10.1128/jvi.11.6.848-855.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Francke B., Ray D. S. Fate of parental phi X174-DNA upon infection of starved thymine-requiring host cells. Virology. 1971 Apr;44(1):168–187. doi: 10.1016/0042-6822(71)90163-2. [DOI] [PubMed] [Google Scholar]
  6. Hirota Y., Ryter A., Jacob F. Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968;33:677–693. doi: 10.1101/sqb.1968.033.01.077. [DOI] [PubMed] [Google Scholar]
  7. Kranias E. G., Dumas L. B. Replication of bacteriophage phiX174 DNA in a temperature-sensitive dnaC mutant of escherichia coli C. J Virol. 1974 Jan;13(1):146–154. doi: 10.1128/jvi.13.1.146-154.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Levine A. J., Sinsheimer R. L. The process of infection with bacteriophage phiX174. XXV. Studies with bacteriophage phiX174 mutants blocked in progeny replicative form DNA synthesis. J Mol Biol. 1969 Feb 14;39(3):619–639. doi: 10.1016/0022-2836(69)90149-1. [DOI] [PubMed] [Google Scholar]
  9. Lindqvist B. H., Sinsheimer R. L. Process of infection with bacteriophage phi-X174. XIV. Studies on macromolecular synthesis during infection with a lysis-defective mutant. J Mol Biol. 1967 Aug 28;28(1):87–94. doi: 10.1016/s0022-2836(67)80079-2. [DOI] [PubMed] [Google Scholar]
  10. Lindqvist B. H., Sinsheimer R. L. The process of infection with bacteriophage phi-X174. XV. Bacteriophage DNA synthesis in abortive infections with a set of conditional lethal mutants. J Mol Biol. 1967 Nov 28;30(1):69–80. doi: 10.1016/0022-2836(67)90244-6. [DOI] [PubMed] [Google Scholar]
  11. Newbold J. E., Sinsheimer R. L. The process of infection with bacteriophage phiX174. XXXII. Early steps in the infection process: attachment, eclipse and DNA penetration. J Mol Biol. 1970 Apr 14;49(1):49–66. doi: 10.1016/0022-2836(70)90375-x. [DOI] [PubMed] [Google Scholar]
  12. Olsen W. L., Staudenbauer W. L., Hofschneider P. H. Replication of bacteriophage M13: specificity of the Escherichia coli dnaB function for replication of double-stranded M13 DNA. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2570–2573. doi: 10.1073/pnas.69.9.2570. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Primrose S. B., Landers W. M., Dowell C. E. Replication of bacteriophage phiX174 in a temperature-sensitive deoxyribonucleic acid host cell. J Virol. 1971 Oct;8(4):594–596. doi: 10.1128/jvi.8.4.594-596.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. SINSHEIMER R. L., STARMAN B., NAGLER C., GUTHRIE S. The process of infection with bacteriophage phi-XI74. I. Evidence for a "replicative form". J Mol Biol. 1962 Mar;4:142–160. doi: 10.1016/s0022-2836(62)80047-3. [DOI] [PubMed] [Google Scholar]
  15. Schekman R., Wickner W., Westergaard O., Brutlag D., Geider K., Bertsch L. L., Kornberg A. Initiation of DNA synthesis: synthesis of phiX174 replicative form requires RNA synthesis resistant to rifampicin. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2691–2695. doi: 10.1073/pnas.69.9.2691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schneck P. K., Staudenbauter W. L., Hofschneider P. H. Replication of bacteriophage M-13. Template specific inhibition of DNA synthesis by nalidixic acid. Eur J Biochem. 1973 Sep 21;38(1):130–136. doi: 10.1111/j.1432-1033.1973.tb03042.x. [DOI] [PubMed] [Google Scholar]
  17. Siccardi A. G., Shapiro B. M. On the process of cellular division in Escherichia coli. IV. Altered protein composition and turnover of the membranes of thermosensitive mutants defective in chromosomal replication. J Mol Biol. 1971 Mar 28;56(3):475–490. doi: 10.1016/0022-2836(71)90395-0. [DOI] [PubMed] [Google Scholar]
  18. Sinsheimer R. L., Knippers R., Komano T. Stages in the replication of bacteriophage phi X174 DNA in vivo. Cold Spring Harb Symp Quant Biol. 1968;33:443–447. doi: 10.1101/sqb.1968.033.01.051. [DOI] [PubMed] [Google Scholar]
  19. Staudenbauer W. L., Hofschneider P. H. Replication of bacteriophage M 13. Mechanism of single-strand DNA synthesis in an escherichia coli mutant thermosensitive in chromosomal DNA replication. Eur J Biochem. 1972 Nov 7;30(3):403–412. doi: 10.1111/j.1432-1033.1972.tb02111.x. [DOI] [PubMed] [Google Scholar]
  20. Steinberg R. A., Denhardt D. T. Inhibition of synthesis of phiX174 DNA in a mutant host thermosensitive for DNA synthesis. J Mol Biol. 1968 Nov 14;37(3):525–528. doi: 10.1016/0022-2836(68)90120-4. [DOI] [PubMed] [Google Scholar]
  21. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]
  22. Wickner W., Brutlag D., Schekman R., Kornberg A. RNA synthesis initiates in vitro conversion of M13 DNA to its replicative form. Proc Natl Acad Sci U S A. 1972 Apr;69(4):965–969. doi: 10.1073/pnas.69.4.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wright M., Wickner S., Hurwitz J. Studies on in vitro DNA synthesis. Isolation of DNA B gene product from Escherichia coli. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3120–3124. doi: 10.1073/pnas.70.11.3120. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES