Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1975 Oct;16(4):775–782. doi: 10.1128/jvi.16.4.775-782.1975

Isolation and partial characterization of single-stranded adenoviral DNA produced during synthesis of adenovirus type 2 DNA.

G Lavelle, C Patch, G Khoury, J Rose
PMCID: PMC354735  PMID: 1165594

Abstract

Single-stranded fragments of adenovirus type 2 DNA were isolated from infected KB cells under conditions which retarded reassociation of complementary sequences but did not denature native viral DNA. Of the total intracellular, virus-specific DNA labeled during a 1-h pulse with tritiated thymidine begining 15 h after infection, about 20% was single stranded when fractionated on hydroxylapatite. This DNA shifted predominantly to the double-stranded fraction on hydroxylapatite during an extended chase incubation, suggesting that it may represent single-stranded DNA in replicating intermediates. Furthermore, the single-stranded DNA annealed nearly equally to both strands of the adenovirus genome. These findings indicate that at least portions of both complementary strands of adenovirus type 2 DNA are exposed as single strands during the period of viral DNA synthesis.

Full text

PDF
780

Selected References

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

  1. Bellett A. J., Younghusband H. B. Replication of the DNA of chick embryo lethal orphan virus. J Mol Biol. 1972 Dec 30;72(3):691–709. doi: 10.1016/0022-2836(72)90185-4. [DOI] [PubMed] [Google Scholar]
  2. Berns K. I., Rose J. A. Evidence for a single-stranded adenovirus-associated virus genome: isolation and separation of complementary single strands. J Virol. 1970 Jun;5(6):693–699. doi: 10.1128/jvi.5.6.693-699.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bourgaux P., Bourgaux-Ramoisy D., Dulbecco R. The replication of the ring-shaped DNA of polyoma virus. I. Identification of the replicative intermediate. Proc Natl Acad Sci U S A. 1969 Oct;64(2):701–708. doi: 10.1073/pnas.64.2.701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Doerfler W., Kleinschmidt A. K. Denaturation pattern of the DNA of adenovirus type 2 as determined by electron microscopy. J Mol Biol. 1970 Jun 28;50(3):579–593. doi: 10.1016/0022-2836(70)90086-0. [DOI] [PubMed] [Google Scholar]
  5. Garon C. F., Berry K. W., Rose J. A. A unique form of terminal redundancy in adenovirus DNA molecules. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2391–2395. doi: 10.1073/pnas.69.9.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gelb L. D., Kohne D. E., Martin M. A. Quantitation of Simian virus 40 sequences in African green monkey, mouse and virus-transformed cell genomes. J Mol Biol. 1971 Apr 14;57(1):129–145. doi: 10.1016/0022-2836(71)90123-9. [DOI] [PubMed] [Google Scholar]
  7. Green M., Piña M., Kimes R., Wensink P. C., MacHattie L. A., Thomas C. A., Jr Adenovirus DNA. I. Molecular weight and conformation. Proc Natl Acad Sci U S A. 1967 May;57(5):1302–1309. doi: 10.1073/pnas.57.5.1302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Horwitz M. S. Intermediates in the synthesis of type 2 adenovirus deoxyribonucleic acid. J Virol. 1971 Nov;8(5):675–683. doi: 10.1128/jvi.8.5.675-683.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Koczot F. J., Carter B. J., Garon C. F., Rose J. A. Self-complementarity of terminal sequences within plus or minus strands of adenovirus-associated virus DNA. Proc Natl Acad Sci U S A. 1973 Jan;70(1):215–219. doi: 10.1073/pnas.70.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Patch C. T., Lewis A. M., Jr, Levine A. S. Evidence for a transcription-control region of Simian virus 40 in the adenovirus 2--Simian virus 40 hybrid, Ad2+ND 1 . Proc Natl Acad Sci U S A. 1972 Nov;69(11):3375–3379. doi: 10.1073/pnas.69.11.3375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Reich P. R., Baum S. G., Rose J. A., Rowe W. P., Weissman S. M. Nucleic acid homology studies of adenovirus type 7-SV40 interactions. Proc Natl Acad Sci U S A. 1966 Feb;55(2):336–341. doi: 10.1073/pnas.55.2.336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Robinson A. J., Younghusband H. B., Bellett A. J. A circula DNA-protein complex from adenoviruses. Virology. 1973 Nov;56(1):54–69. doi: 10.1016/0042-6822(73)90287-0. [DOI] [PubMed] [Google Scholar]
  13. Rose J. A., Berns K. I., Hoggan M. D., Koczot F. J. Evidence for a single-stranded adenovirus-associated virus genome: formation of a DNA density hybrid on release of viral DNA. Proc Natl Acad Sci U S A. 1969 Nov;64(3):863–869. doi: 10.1073/pnas.64.3.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rose J. A., Hoggan M. D., Shatkin A. J. Nucleic acid from an adeno-associated virus: chemical and physical studies. Proc Natl Acad Sci U S A. 1966 Jul;56(1):86–92. doi: 10.1073/pnas.56.1.86. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rose J. A., Koczot F. Adenovirus-associated virus multiplication. VI. Base compostion of the deoxyribonucleic acid strand species and strand-specific in vivo transcription. J Virol. 1971 Nov;8(5):771–777. doi: 10.1128/jvi.8.5.771-777.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rose J. A., Koczot F. Adenovirus-associated virus multiplication. VII. Helper requirement for viral deoxyribonucleic acid and ribonucleic acid synthesis. J Virol. 1972 Jul;10(1):1–8. doi: 10.1128/jvi.10.1.1-8.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sussenbach J. S., van der Vliet P. C., Ellens D. J., Jansz H. S. Linear intermediates in the replication of adenovirus DNA. Nat New Biol. 1972 Sep 13;239(89):47–49. [PubMed] [Google Scholar]
  18. Sussenbach J. S., van der Vliet P. C. Studies on the mechanism of replication of adenovirus DNA. I. The effect of hydroxyurea. Virology. 1973 Jul;54(1):299–303. doi: 10.1016/0042-6822(73)90142-6. [DOI] [PubMed] [Google Scholar]
  19. Tolun A., Pettersson U. Termination sites for adenovirus type 2 DNA replication. J Virol. 1975 Oct;16(4):759–766. doi: 10.1128/jvi.16.4.759-766.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Warmaar S. O., Cohen J. A. A quantitative assay for DNA-DNA hybrids using membrane filters. Biochem Biophys Res Commun. 1966 Aug 23;24(4):554–558. doi: 10.1016/0006-291x(66)90356-1. [DOI] [PubMed] [Google Scholar]
  21. Wolfson J., Dressler D. Adenovirus-2 DNA contains an inverted terminal repetition. Proc Natl Acad Sci U S A. 1972 Oct;69(10):3054–3057. doi: 10.1073/pnas.69.10.3054. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. van der Eb A. J., van Kesteren L. W., van Bruggen E. F. Structural properties of adenovirus DNA's. Biochim Biophys Acta. 1969 Jun 17;182(2):530–541. doi: 10.1016/0005-2787(69)90205-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES