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. 1974 Oct;14(4):872–877. doi: 10.1128/jvi.14.4.872-877.1974

Location of the 5-Methylcytosine Group on the Bacteriophage φX174 Genome

Amy Shiu Lee 1, Robert L Sinsheimer 1
PMCID: PMC355593  PMID: 4609216

Abstract

Bacteriophage φX174 DNA was labeled in vivo with [methyl-3H]methionine. The methyl-labeled progeny DNA was extracted from purified bacteriophage φX174 particles and was used as template for in vitro synthesis of the complementary strand in the presence of the nucleoside triphosphates and Escherichia coli polymerase I. The resultant replicative form DNA was then cleaved, in separate experiments, with restriction endonucleases from Haemophilus influenzae and H. aegyptius. The DNA fragments were analyzed by polyacrylamide gel electrophoresis. It is concluded that the single methylcytosine in the viral DNA is located in a specific region of the φX174 genome, very likely in gene H.

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Selected References

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

  1. Arber W., Linn S. DNA modification and restriction. Annu Rev Biochem. 1969;38:467–500. doi: 10.1146/annurev.bi.38.070169.002343. [DOI] [PubMed] [Google Scholar]
  2. Benbow R. M., Zuccarelli A. J., Davis G. C., Sinsheimer R. L. Genetic recombination in bacteriophage phi chi 174. J Virol. 1974 Apr;13(4):898–907. doi: 10.1128/jvi.13.4.898-907.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Billen D. Methylation of the bacterial chromosome: an event at the "replication point"? J Mol Biol. 1968 Feb 14;31(3):477–486. doi: 10.1016/0022-2836(68)90422-1. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Edgell M. H., Hutchison C. A., 3rd, Sclair M. Specific endonuclease R fragments of bacteriophage phiX174 deoxyribonucleic acid. J Virol. 1972 Apr;9(4):574–582. doi: 10.1128/jvi.9.4.574-582.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gilbert W., Dressler D. DNA replication: the rolling circle model. Cold Spring Harb Symp Quant Biol. 1968;33:473–484. doi: 10.1101/sqb.1968.033.01.055. [DOI] [PubMed] [Google Scholar]
  7. Johnson P. H., Lee A. S., Sinsheimer R. L. Production of specific fragments of phi X174 replicative form DNA by a restriction enzyme from Haemophilus parainfluenzae, endonuclease HP. J Virol. 1973 Apr;11(4):596–599. doi: 10.1128/jvi.11.4.596-599.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Johnson P. H., Sinsheimer R. L. Structure of an intermediate in the replication of bacteriophage phiX174 deoxyribonucleic acid: the initiation site for DNA replication. J Mol Biol. 1974 Feb 15;83(1):47–61. doi: 10.1016/0022-2836(74)90423-9. [DOI] [PubMed] [Google Scholar]
  9. Kelly T. J., Jr, Smith H. O. A restriction enzyme from Hemophilus influenzae. II. J Mol Biol. 1970 Jul 28;51(2):393–409. doi: 10.1016/0022-2836(70)90150-6. [DOI] [PubMed] [Google Scholar]
  10. Lark C. Effect of the methionine analogs, ethionine and norleucine, on DNA synthesis in Escherichia coli 15T. J Mol Biol. 1968 Feb 14;31(3):401–414. doi: 10.1016/0022-2836(68)90417-8. [DOI] [PubMed] [Google Scholar]
  11. Lark C. Studies on the in vivo methylation of DNA in Escherichia coli 15T. J Mol Biol. 1968 Feb 14;31(3):389–399. doi: 10.1016/0022-2836(68)90416-6. [DOI] [PubMed] [Google Scholar]
  12. Lee A. S., Sinsheimer R. L. A cleavage map of bacteriophage phiX174 genome. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2882–2886. doi: 10.1073/pnas.71.7.2882. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lee A. S., Sinsheimer R. L. A continuous electroelution method for the recovery of DNA restriction enzyme fragments. Anal Biochem. 1974 Aug;60(2):640–644. doi: 10.1016/0003-2697(74)90279-6. [DOI] [PubMed] [Google Scholar]
  14. Middleton J. H., Edgell M. H., Hutchison C. A., 3rd Specific fragments of phi X174 deoxyribonucleic acid produced by a restriction enzyme from Haemophilus aegyptius, endonuclease Z. J Virol. 1972 Jul;10(1):42–50. doi: 10.1128/jvi.10.1.42-50.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Razin A., Sedat J. W., Sinsheimer R. L. In vivo methylation of replicating bacteriophage phi chi174 DNA. J Mol Biol. 1973 Aug 15;78(3):417–425. doi: 10.1016/0022-2836(73)90465-8. [DOI] [PubMed] [Google Scholar]
  16. Razin A., Sedat J. W., Sinsheimer R. L. Structure of the DNA of bacteriophage phiX174. VII. Methylation. J Mol Biol. 1970 Oct 28;53(2):251–259. doi: 10.1016/0022-2836(70)90298-6. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Scarano E., Iaccarino M., Grippo P., Parisi E. The heterogeneity of thymine methyl group origin in DNA pyrimidine isostichs of developing sea urchin embryos. Proc Natl Acad Sci U S A. 1967 May;57(5):1394–1400. doi: 10.1073/pnas.57.5.1394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Smith H. O., Wilcox K. W. A restriction enzyme from Hemophilus influenzae. I. Purification and general properties. J Mol Biol. 1970 Jul 28;51(2):379–391. doi: 10.1016/0022-2836(70)90149-x. [DOI] [PubMed] [Google Scholar]
  20. Weisbeek P. J., van de Pol J. H. Bological activity of phi X174 replicative form DNA fragments. Biochim Biophys Acta. 1970 Dec 14;224(2):328–338. [PubMed] [Google Scholar]

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