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. 1972 Jul;10(1):23–31. doi: 10.1128/jvi.10.1.23-31.1972

Role of Subunits of 60 to 70S Avian Tumor Virus Ribonucleic Acid in Its Template Activity for the Viral Deoxyribonucleic Acid Polymerase

Eli Canaani 1, Peter Duesberg 1
PMCID: PMC356421  PMID: 4339194

Abstract

Heating the 60 to 70S ribonucleic acid (RNA) of Rous sarcoma virus (RSV) destroys both its subunit structure and its high template activity for RSV deoxyribonucleic acid (DNA) polymerase. In comparative analyses, it was found that the template activity of the RNA has a thermal transition of 70 C, whereas the 60 to 70S structure dissociates into 30 to 40S and several distinct small subunits with a Tm of 55 C. Analysis by velocity sedimentation and isopycnic centrifugation of the primary DNA product obtained by incubation of 60 to 70S RSV RNA with RSV DNA polymerase indicated that most, but perhaps not all, DNA was linked to small (<10S) RSV RNA primer. Sixty percent of the high template activity of 60 to 70S RSV RNA lost after heat dissociation could be recovered by incubation of the total RNA under annealing conditions. The template activity of purified 30 to 40S subunits isolated from 60 to 70S RSV RNA was not enhanced significantly by annealing. However, in the presence of small (<10S) subunits also isolated from 60 to 70S RNA, the template activity of 30 to 40S RNA subunits was increased to the same level as that of reannealed total 60 to 70S RNA. It was concluded that neither the 30 to 40S subunits nor most of the 4S subunits of 60 to 70S RSV RNA contribute much as primers to the template activity of 60 to 70S RSV RNA. The predominant primer molecule appears to be a minor component of the <10S subunit fraction of 60 to 70S RSV RNA. Its electrophoretic mobility is similar to, and its dissociation temperature from 60 to 70S RSV RNA is higher than that of the bulk of 60 to 70S RSV RNA-associated 4S RNA. The role of primers in DNA synthesis by RSV DNA polymerase is discussed.

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Selected References

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

  1. Bader J. P., Steck T. L. Analysis of the ribonucleic acid of murine leukemia virus. J Virol. 1969 Oct;4(4):454–459. doi: 10.1128/jvi.4.4.454-459.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baltimore D., Smoler D. Primer requirement and template specificity of the DNA polymerase of RNA tumor viruses. Proc Natl Acad Sci U S A. 1971 Jul;68(7):1507–1511. doi: 10.1073/pnas.68.7.1507. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boedtker H. Dependence of the sedimentation coefficient on molecular weight of RNA after reaction with formaldehyde. J Mol Biol. 1968 Jul 14;35(1):61–70. doi: 10.1016/s0022-2836(68)80036-1. [DOI] [PubMed] [Google Scholar]
  4. Brutlag D., Schekman R., Kornberg A. A possible role for RNA polymerase in the initiation of M13 DNA synthesis. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2826–2829. doi: 10.1073/pnas.68.11.2826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Duesberg P. H., Canaani E. Complementarity between Rous sarcoma virus (RSV) RNA and the in vitro-synthesized DNA of the virus-associated DNA polymerase. Virology. 1970 Nov;42(3):783–788. doi: 10.1016/0042-6822(70)90325-9. [DOI] [PubMed] [Google Scholar]
  6. Duesberg P. H., Cardiff R. D. Structural relationships between the RNA of mammary tumor virus and those of other RNA tumor viruses. Virology. 1968 Dec;36(4):696–700. doi: 10.1016/0042-6822(68)90206-7. [DOI] [PubMed] [Google Scholar]
  7. Duesberg P. H. On the structure of RNA tumor viruses. Curr Top Microbiol Immunol. 1970;51:78–104. [PubMed] [Google Scholar]
  8. Duesberg P. H. Physical properties of Rous Sarcoma Virus RNA. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1511–1518. doi: 10.1073/pnas.60.4.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Duesberg P. H., Vogt P. K. Differences between the ribonucleic acids of transforming and nontransforming avian tumor viruses. Proc Natl Acad Sci U S A. 1970 Dec;67(4):1673–1680. doi: 10.1073/pnas.67.4.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Duesberg P., Helm K. V., Canaani E. Comparative properties of RNA and DNA templates for the DNA polymerase of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2505–2509. doi: 10.1073/pnas.68.10.2505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Duesberg P., Helm K. V., Canaani E. Properties of a soluble DNA polymerase isolated from Rous sarcoma virus. Proc Natl Acad Sci U S A. 1971 Apr;68(4):747–751. doi: 10.1073/pnas.68.4.747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Erikson E., Erikson R. L. Association of 4S ribonucleic acid with oncornavirus ribonucleic acids. J Virol. 1971 Aug;8(2):254–256. doi: 10.1128/jvi.8.2.254-256.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Erikson R. L. Studies on the RNA from avian myeloblastosis virus. Virology. 1969 Jan;37(1):124–131. doi: 10.1016/0042-6822(69)90313-4. [DOI] [PubMed] [Google Scholar]
  14. HURWITZ J. The enzymatic incorporation of ribonucleotides into polydeoxynucletide material. J Biol Chem. 1959 Sep;234:2351–2358. [PubMed] [Google Scholar]
  15. Hurwitz J., Leis J. P. RNA-dependent DNA polymerase activity of RNA tumor viruses. I. Directing influence of DNA in the reaction. J Virol. 1972 Jan;9(1):116–129. doi: 10.1128/jvi.9.1.116-129.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lark K. G. Evidence for the direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol. 1972 Feb 28;64(1):47–60. doi: 10.1016/0022-2836(72)90320-8. [DOI] [PubMed] [Google Scholar]
  17. Leis J. P., Hurwitz J. RNA-dependent DNA polymerase activity of RNA tumor viruses. II. Directing influence of RNA in the reaction. J Virol. 1972 Jan;9(1):130–142. doi: 10.1128/jvi.9.1.130-142.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Montagnier L., Goldé A., Vigier P. A possible subunit structure of Rous sarcoma virus RNA. J Gen Virol. 1969 Apr;4(3):449–452. doi: 10.1099/0022-1317-4-3-449. [DOI] [PubMed] [Google Scholar]
  19. Verma I. M., Meuth N. L., Bromfeld E., Manly K. F., Baltimore D. Covalently linked RNA-DNA molecule as initial product of RNA tumour virus DNA polymerase. Nat New Biol. 1971 Sep 29;233(39):131–134. doi: 10.1038/newbio233131a0. [DOI] [PubMed] [Google Scholar]
  20. Zimmerman S. B., Sandeen D. The ribonuclease activity of crystallized pancreatic deoxyribonuclease. Anal Biochem. 1966 Feb;14(2):269–277. doi: 10.1016/0003-2697(66)90137-0. [DOI] [PubMed] [Google Scholar]

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