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. 1967 Jun;1(3):514–522. doi: 10.1128/jvi.1.3.514-522.1967

Replication of Bacteriophage Ribonucleic Acid: Some Properties of Native and Denatured Replicative Intermediate

Richard M Franklin 1
PMCID: PMC375269  PMID: 5623970

Abstract

Purified replicative form (RF) and replicative intermediate (RI) prepared from Escherichia coli cells infected with the ribonucleic acid (RNA) bacteriophage R17 were denatured with dimethyl sulfoxide at 37 C or in aqueous solvents of low ionic strength at 97 C. Denaturation was demonstrated for RF and RI by an increase in specific infectivity and a striking change in the hyperchromicity curves after treatment. RI denaturation was also demonstrated by a shift in the buoyant density in Cs2SO4 from 1.619 to the buoyant density of single-stranded R17 RNA (1.627). Analysis of the denatured RI hyperchromicity curves and the equilibrium distributions of denatured RI in Cs2SO4 gradients revealed, however, a residual double-stranded component. Velocity sedimentation of denatured RI was performed, and the weight distribution of S values was calculated. From the known relation between molecular weight and S values, it was possible to transform the weight distribution into a number distribution of chain lengths. This distribution was compared with that predicted from the steady-state hypothesis for RI. Deviations from the predicted distribution may be due to the residual double-stranded component.

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Selected References

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

  1. AMMANN J., DELIUS H., HOFSCHNEIDER P. H. ISOLATION AND PROPERTIES OF AN INTACT PHAGE-SPECIFIC REPLICATIVE FORM OF RNA PHAGE M12. J Mol Biol. 1964 Dec;10:557–561. doi: 10.1016/s0022-2836(64)80079-6. [DOI] [PubMed] [Google Scholar]
  2. Doty P., Boedtker H., Fresco J. R., Haselkorn R., Litt M. SECONDARY STRUCTURE IN RIBONUCLEIC ACIDS. Proc Natl Acad Sci U S A. 1959 Apr;45(4):482–499. doi: 10.1073/pnas.45.4.482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Erikson R. L. Fractionation of viral-specific ribonucleic acid by cesium sulfate equilibrium density-gradient centrifugation. J Mol Biol. 1966 Jul;18(2):372–381. doi: 10.1016/s0022-2836(66)80254-1. [DOI] [PubMed] [Google Scholar]
  4. Erikson R. L., Franklin R. M. Symposium on replication of viral nucleic acids. I. Formation and properties of a replicative intermediate in the biosynthesis of viral ribonucleic acid. Bacteriol Rev. 1966 Jun;30(2):267–278. doi: 10.1128/br.30.2.267-278.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Francke B., Hofschneider P. H. Infectious nucleic acids of e. Coli bacteriophages, ix. Sedimentation constants and strand integrity of infectious m12 phage replicative-form RNA. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1883–1890. doi: 10.1073/pnas.56.6.1883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Franklin R. M., Granboulan N. Ultrastructure of Escherichia coli cells infected with bacteriophage R17. J Bacteriol. 1966 Feb;91(2):834–848. doi: 10.1128/jb.91.2.834-848.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Franklin R. M. Purification and properties of the replicative intermediate of the RNA bacteriophage R17. Proc Natl Acad Sci U S A. 1966 Jun;55(6):1504–1511. doi: 10.1073/pnas.55.6.1504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. GESTELAND R. F., BOEDTKER H. SOME PHYSICAL PROPERTIES OF BACTERIOPHAGE R17 AND ITS RIBONUCLEIC ACID. J Mol Biol. 1964 Apr;8:496–507. doi: 10.1016/s0022-2836(64)80007-3. [DOI] [PubMed] [Google Scholar]
  9. Gomatos P. J., Tamm I. THE SECONDARY STRUCTURE OF REOVIRUS RNA. Proc Natl Acad Sci U S A. 1963 May;49(5):707–714. doi: 10.1073/pnas.49.5.707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. KAERNER H. C., HOFFMANN-BERLING H. SYNTHESIS OF DOUBLE-STRANDED RNA IN RNA-PHAGE INFECTED E. COLI CELLS. Nature. 1964 Jun 6;202:1012–1013. doi: 10.1038/2021012a0. [DOI] [PubMed] [Google Scholar]
  11. Katz L., Penman S. The solvent denaturation of double-stranded RNA from poliovirus infected HeLa cells. Biochem Biophys Res Commun. 1966 May 25;23(4):557–560. doi: 10.1016/0006-291x(66)90765-0. [DOI] [PubMed] [Google Scholar]
  12. MONTAGNIER L., SANDERS F. K. REPLICATIVE FORM OF ENCEPHALOMYOCARDITIS VIRUS RIBONUCLEIC ACID. Nature. 1963 Aug 17;199:664–667. doi: 10.1038/199664a0. [DOI] [PubMed] [Google Scholar]
  13. SCHUMAKER V. N., SCHACHMAN H. K. Ultracentrifugal analysis of dilute solutions. Biochim Biophys Acta. 1957 Mar;23(3):628–639. doi: 10.1016/0006-3002(57)90386-4. [DOI] [PubMed] [Google Scholar]
  14. SINHA N. K., FUJIMURA R. K., KAESBERG P. RIBONUCLEASE DIGESTION OF R17 VIRAL RNA. J Mol Biol. 1965 Jan;11:84–89. doi: 10.1016/s0022-2836(65)80173-5. [DOI] [PubMed] [Google Scholar]
  15. THOMAS C. A., Jr, BERNS K. I. The physical characterization of DNA molecules released from T2 and T4 bacteriophage. J Mol Biol. 1961 Jun;3:277–288. doi: 10.1016/s0022-2836(61)80069-7. [DOI] [PubMed] [Google Scholar]
  16. VINOGRAD J., HEARST J. E. Equilibrium sedimentation of macromolecules and viruses in a density gradient. Fortschr Chem Org Naturst. 1962;20:373–422. [PubMed] [Google Scholar]
  17. Vasquez C., Granboulan N., Franklin R. M. Structure of the ribonucleic acid bacteriophage R17. J Bacteriol. 1966 Dec;92(6):1779–1786. doi: 10.1128/jb.92.6.1779-1786.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]

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