Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1975 Oct;72(10):3907–3911. doi: 10.1073/pnas.72.10.3907

phiX174 DNA-dependent DNA synthesis in vitro: requirement for P1 ban protein in dnaB mutant extracts of Escherichia coli.

H Schuster, M Mikolajczyk, J Rohrschneider, B Geschke
PMCID: PMC433105  PMID: 670

Abstract

Ammonium sulfate fractionation of crude extracts of E. coli yields a soluble enzyme fraction (about 25-fold purification) that catalyzes the conversion of phiX174 single-stranded DNA to duplex DNA. The reaction is rifampicin-resistant, requires single-stranded DNA, Mg++, deoxynucleoside triphosphates, and ATP, and is stimulated by KCl. Such soluble enzyme fractions were prepared from E. coli strains carrying the prophage mutant P1bac, in which the viral dnaB analog (ban) protein is expressed constitutively, or P1bacban, in which the expression of ban protein is prevented. DNA-synthesizing activity of ban protein containing fractions from wild-type or dnaB(P1bac) lysogens was more temperature-resistant than that from E. coli containing only wild-type dnaB protein, whereas that from dnaB(P1bacban) lysogens of dnaB cells was extremely thermolabile. It is suggested that the temperature-resistant DNA synthesis with fractions from P1bac lysogens is mediated by the P1 ban protein.

Full text

PDF

Selected References

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

  1. Beyersmann D., Schuster H. DNA synthesis in P1 infected E. coli mutants temperature-sensitive in DNA replication. Mol Gen Genet. 1972;114(2):173–176. doi: 10.1007/BF00332788. [DOI] [PubMed] [Google Scholar]
  2. D'Ari R., Jaffé-Brachet A., Touati-Schwartz D., Yarmolinsky M. B. A dnaB analog specified by bacteriophage P1. J Mol Biol. 1975 May 25;94(3):341–366. doi: 10.1016/0022-2836(75)90207-7. [DOI] [PubMed] [Google Scholar]
  3. Hutchison C. A., 3rd, Sinsheimer R. L. The process of infection with bacteriophage phi-X174. X. Mutations in a phi-X Lysis gene. J Mol Biol. 1966 Jul;18(3):429–447. doi: 10.1016/s0022-2836(66)80035-9. [DOI] [PubMed] [Google Scholar]
  4. KONDO E., MITSUHASHI S. DRUG RESISTANCE OF ENTERIC BACTERIA. IV. ACTIVE TRANSDUCING BACTERIOPHAGE P1 CM PRODUCED BY THE COMBINATION OF R FACTOR WITH BACTERIOPHAGE P1. J Bacteriol. 1964 Nov;88:1266–1276. doi: 10.1128/jb.88.5.1266-1276.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Koiyama M., Milewski E. Isolation of a complementing activity for a dna-B mutant. Biochem Biophys Res Commun. 1973 Aug 6;53(3):858–868. doi: 10.1016/0006-291x(73)90172-1. [DOI] [PubMed] [Google Scholar]
  6. Lanka E., Schuster H. Replication of bacteriophages in Escherichia coli mutants thermosensitive in DNA synthesis. Mol Gen Genet. 1970;106(3):274–285. doi: 10.1007/BF00340386. [DOI] [PubMed] [Google Scholar]
  7. Mikolajczyk M., Schuster H. The fate of newly made DNA in Escherichia coli mutants thermosensitive in DNA synthesis. Mol Gen Genet. 1971;110(3):272–288. doi: 10.1007/BF00337839. [DOI] [PubMed] [Google Scholar]
  8. Ogawa T. Analysis of dnaB function of Escherichia coli K12 and the dnaB-like function of P1 prophage. J Mol Biol. 1975 May 25;94(3):327–340. doi: 10.1016/0022-2836(75)90206-5. [DOI] [PubMed] [Google Scholar]
  9. Schekman R., Weiner A., Kornberg A. Multienzyme systems of DNA replication. Science. 1974 Dec 13;186(4168):987–993. doi: 10.1126/science.186.4168.987. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Wechsler J. A., Nüsslein V., Otto B., Klein A., Bonhoeffer F., Herrmann R., Gloger L., Schaller H. Isolation and characterization of thermosensitive Escherichia coli mutants defective in deoxyribonucleic acid replication. J Bacteriol. 1973 Mar;113(3):1381–1388. doi: 10.1128/jb.113.3.1381-1388.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Wickner S., Hurwitz J. Conversion of phiX174 viral DNA to double-stranded form by purified Escherichia coli proteins. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4120–4124. doi: 10.1073/pnas.71.10.4120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. 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 Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES