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. 1976 Apr;18(1):245–259. doi: 10.1128/jvi.18.1.245-259.1976

Multiple origins and circular structures in replicating T5 bacteriophage DNA.

G J Bourguignon, T K Sweeney, H Delius
PMCID: PMC515544  PMID: 1255872

Abstract

Replicating T5 phage DNA was gently isolated using NaI density gradient centrifugation and examined by electron microscopy. At the beginning of phage DNA synthesis, linear unit-length T5 DNA molecules containing from one to four replicating "eye-loops" were consistently observed. Replication in these molecules was found to proceed bidirectionally from multiple, internal origins. A primary origin of replication is located near the center of the T5 genome, which does not coincide with the location of any of the nicks (single-strand breaks) found in mature T5 DNA. The initiation of replication at the various origins within an individual molecule does not appear to follow any definite temporal sequence. At later times in the infection, we have observed a significant number of circular T5 DNA molecules-both replicating and nonreplicating-whose average circumference is approximately the length of mature T5 DNA minus the terminal redundancy. The replicating circular molecules appear to be either in a theta configuration, a sigma configuration with the tails all being less than the length of the circle, or a combination of theta and sigma forms.

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Selected References

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

  1. Anet R., Strayer D. R. Sodium iodide density gradients for the preparative buoyant density separations of DNA mixtures. Biochem Biophys Res Commun. 1969 Sep 24;37(1):52–58. doi: 10.1016/0006-291x(69)90879-1. [DOI] [PubMed] [Google Scholar]
  2. Benzinger R., Scheible P. Transfection of Escherichia coli spheroplasts. IV. Transfection of rec+ and rec minus spheroplasts by native, denatured, and renatured T5 bacteriophage DNA after repair of single-strand breaks by polynucleotide ligase. J Virol. 1974 May;13(5):960–966. doi: 10.1128/jvi.13.5.960-966.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bernstein H., Bernstein C. Circular and branched circular concatenates as possible intermediates in bacteriophage T4 DNA replication. J Mol Biol. 1973 Jul 5;77(3):355–361. doi: 10.1016/0022-2836(73)90443-9. [DOI] [PubMed] [Google Scholar]
  4. Blumenthal A. B., Kriegstein H. J., Hogness D. S. The units of DNA replication in Drosophila melanogaster chromosomes. Cold Spring Harb Symp Quant Biol. 1974;38:205–223. doi: 10.1101/sqb.1974.038.01.024. [DOI] [PubMed] [Google Scholar]
  5. Bujard H., Hendrickson H. E. Structure and function of the genome of coliphage T5. 1. The physical structure of the chromosome of T5 + . Eur J Biochem. 1973 Mar 15;33(3):517–528. doi: 10.1111/j.1432-1033.1973.tb02711.x. [DOI] [PubMed] [Google Scholar]
  6. Carrington J. M., Lunt M. R. Studies on the replication of bacteriophage T5. J Gen Virol. 1973 Feb;18(2):91–109. doi: 10.1099/0022-1317-18-2-91. [DOI] [PubMed] [Google Scholar]
  7. De Kloet S. R., Andrean B. A. Buoyant density gradient centrifugation of RNA and DNA in alkali iodide solutions. Biochim Biophys Acta. 1971 Nov 19;247(4):519–527. doi: 10.1016/0005-2787(71)90688-5. [DOI] [PubMed] [Google Scholar]
  8. De Waard A., Paul A. V., Lehman I. R. The structural gene for deoxyribonucleic acid polymerase in bacteriophages T4 and T5. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1241–1248. doi: 10.1073/pnas.54.4.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Delius H., Howe C., Kozinski A. W. Structure of the replicating DNA from bacteriophage T4. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3049–3053. doi: 10.1073/pnas.68.12.3049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Delius H., Mantell N. J., Alberts B. Characterization by electron microscopy of the complex formed between T4 bacteriophage gene 32-protein and DNA. J Mol Biol. 1972 Jun 28;67(3):341–350. doi: 10.1016/0022-2836(72)90454-8. [DOI] [PubMed] [Google Scholar]
  11. Doermann A. H. T4 and the rolling circle model of replication. Annu Rev Genet. 1973;7:325–341. doi: 10.1146/annurev.ge.07.120173.001545. [DOI] [PubMed] [Google Scholar]
  12. Dressler D., Wolfson J., Magazin M. Initiation and reinitiation of DNA synthesis during replication of bacteriophage T7. Proc Natl Acad Sci U S A. 1972 Apr;69(4):998–1002. doi: 10.1073/pnas.69.4.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dressler D., Wolfson J. The rolling circle for phi X DNA replication. 3. Synthesis of supercoiled duplex rings. Proc Natl Acad Sci U S A. 1970 Sep;67(1):456–463. doi: 10.1073/pnas.67.1.456. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hayward G. S. Unique double-stranded fragments of bacteriophage T5 DNA resulting from preferential shear-induced breakage at nicks. Proc Natl Acad Sci U S A. 1974 May;71(5):2108–2112. doi: 10.1073/pnas.71.5.2108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hendrickson H. E., McCorquodale D. J. Genetic and physiological studies of bacteriophage T5. 3. Patterns of deoxyribonucleic acid synthesis induced by mutants of T5 and the identification of genes influencing the appearance of phage-induced dihydrofolate reductase and deoxyribonuclease. J Virol. 1972 Jun;9(6):981–989. doi: 10.1128/jvi.9.6.981-989.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Herman R. C., Moyer R. W. In vivo repair of the single-strand interruptions contained in bacteriophage T5 DNA. Proc Natl Acad Sci U S A. 1974 Mar;71(3):680–684. doi: 10.1073/pnas.71.3.680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hourcade D., Dressler D., Wolfson J. The amplification of ribosomal RNA genes involves a rolling circle intermediate. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2926–2930. doi: 10.1073/pnas.70.10.2926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Howe C. C., Buckley P. J., Carlson K. M., Kozinski A. W. Multiple and specific initiation of T4 DNA replication. J Virol. 1973 Jul;12(1):130–148. doi: 10.1128/jvi.12.1.130-148.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Huberman J. A., Riggs A. D. On the mechanism of DNA replication in mammalian chromosomes. J Mol Biol. 1968 Mar 14;32(2):327–341. doi: 10.1016/0022-2836(68)90013-2. [DOI] [PubMed] [Google Scholar]
  20. Klein A., Bonhoeffer F. DNA replication. Annu Rev Biochem. 1972;41(10):301–332. doi: 10.1146/annurev.bi.41.070172.001505. [DOI] [PubMed] [Google Scholar]
  21. Knippers R., Razin A., Davis R., Sinsheimer R. L. The process of infection with Bacteriophage phi-X174. XXIX. In vivo studies on the synthesis of the single-stranded DNA of progeny phi-X174 bacteriophage. J Mol Biol. 1969 Oct 28;45(2):237–263. doi: 10.1016/0022-2836(69)90103-x. [DOI] [PubMed] [Google Scholar]
  22. Kozinski A. W., Doermann A. H. Repetitive DNA replication of the incomplete genomes of phage T4 petite particles. Proc Natl Acad Sci U S A. 1975 May;72(5):1734–1738. doi: 10.1073/pnas.72.5.1734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lang D. Molecular weights of coliphages and coliphage DNA. 3. Contour length and molecular weight of DNA from bacteriophages T4, T5 and T7, and from bovine papilloma virus. J Mol Biol. 1970 Dec 28;54(3):557–565. doi: 10.1016/0022-2836(70)90126-9. [DOI] [PubMed] [Google Scholar]
  24. Levine A. J., Kang H. S., Billheimer F. E. DNA replication in SV40 infected cells. I. Analysis of replicating SV40 DNA. J Mol Biol. 1970 Jun 14;50(2):549–568. doi: 10.1016/0022-2836(70)90211-1. [DOI] [PubMed] [Google Scholar]
  25. Newlon C. S., Petes T. D., Hereford L. M., Fangman W. L. Replication of yeast chromosomal DNA. Nature. 1974 Jan 4;247(5435):32–35. doi: 10.1038/247032a0. [DOI] [PubMed] [Google Scholar]
  26. Pettersson U., Mulder C., Deluis H., Sharp P. A. Cleavage of adenovirus type 2 DNA into six unique fragments by endonuclease R-RI. Proc Natl Acad Sci U S A. 1973 Jan;70(1):200–204. doi: 10.1073/pnas.70.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rhoades M., Rhoades E. A. Terminal repetition in the DNA of bacteriophage T5. J Mol Biol. 1972 Aug 21;69(2):187–200. doi: 10.1016/0022-2836(72)90224-0. [DOI] [PubMed] [Google Scholar]
  28. Robberson D. L., Clayton D. A. Replication of mitochondrial DNA in mouse L cells and their thymidine kinase - derivatives: displacement replication on a covalently-closed circular template. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3810–3814. doi: 10.1073/pnas.69.12.3810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rochaix J. D., Bird A., Barkken A. Ribosomal RNA gene amplification by rolling circles. J Mol Biol. 1974 Aug 15;87(3):473–487. doi: 10.1016/0022-2836(74)90098-9. [DOI] [PubMed] [Google Scholar]
  30. Rosenkranz H. S. RNA in coliphage T5. Nature. 1973 Mar 30;242(5396):327–329. doi: 10.1038/242327a0. [DOI] [PubMed] [Google Scholar]
  31. Scheible P. P., Rhoades M. Heteroduplex mapping of heat-resistant deletion mutants of bacteriophage t5. J Virol. 1975 May;15(5):1276–1280. doi: 10.1128/jvi.15.5.1276-1280.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Smith M. G., Burton K. Fractionation of deoxyribonucleic acid from phage-infected bacteria. Biochem J. 1966 Jan;98(1):229–241. doi: 10.1042/bj0980229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Smith M. G., Skalka A. Some properties of DNA from phage-infected bacteria. J Gen Physiol. 1966 Jul;49(6):127–142. doi: 10.1085/jgp.49.6.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Studier F. W. The genetics and physiology of bacteriophage T7. Virology. 1969 Nov;39(3):562–574. doi: 10.1016/0042-6822(69)90104-4. [DOI] [PubMed] [Google Scholar]
  35. Warner H. R., Drong R. F., Berget S. M. Early events after infection of Escherichia coli by bacteriophage T5. Induction of a 5'-nucleotidase activity and excretion of free bases. J Virol. 1975 Feb;15(2):273–280. doi: 10.1128/jvi.15.2.273-280.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wolfson J., Dressler D., Magazin M. Bacteriophage T7 DNA replication: a linear replicating intermediate (gradient centrifugation-electron microscopy-E. coli-DNA partial denaturation). Proc Natl Acad Sci U S A. 1972 Feb;69(2):499–504. doi: 10.1073/pnas.69.2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Zweig M., Rosenkranz H. S., Morgan C. Development of coliphage T5: ultrastructural and biochemical studies. J Virol. 1972 Mar;9(3):526–543. doi: 10.1128/jvi.9.3.526-543.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. van der Eb A. J. Intermediates in type 5 adenovirus DNA replication. Virology. 1973 Jan;51(1):11–23. doi: 10.1016/0042-6822(73)90361-9. [DOI] [PubMed] [Google Scholar]

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