Manly, Kenneth F. and Smoler, Donna F. and Bromfeld, Esther and Baltimore, David (1971) Forms of Deoxyribonucleic Acid Produced by Virions of the Ribonucleic Acid Tumor Viruses. Journal of Virology, 7 (1). pp. 106-111. ISSN 0022-538X. http://resolver.caltech.edu/CaltechAUTHORS:MANjvir71
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The in vitro product of mouse leukemia virus deoxyribonucleic acid (DNA) polymerase can be separated into two fractions by sedimentation in sucrose gradients. These two fractions were analyzed for their content of single-stranded DNA, double-stranded DNA, and DNA-ribonucleic acid (RNA) hybrid by (i) digestion with enzymes of known specificity and (ii) equilibrium centrifugation in Cs2SO4 gradients. The major fraction early in the reaction contained equal amounts of single-stranded DNA and DNA-RNA hybrid and little double-stranded DNA. The major fraction after extensive synthesis contained equal amounts of single-and double-stranded DNA and little hybrid. In the presence of actinomycin D, the predominant product was single-stranded DNA. To account for these various forms of DNA, we postulate the following model: the first DNA synthesis occurs in a replicative complex containing growing DNA molecules attached to an RNA molecule. Each DNA molecule is displaced as single-stranded DNA by the synthesis of the following DNA strand, and the single-stranded DNA is copied to form double-stranded DNA either before or after release of the single strand from the RNA. Actinomycin blocks this conversion of single-to double-stranded DNA.
|Additional Information:||Copyright © 1971 American Society for Microbiology. Received for publication 2 November 1970 We thank C. A. Thomas for the supply of endonuclease which made these experiments possible, V. Vogt for the RNA polymerase used in the synthesis of DNA-RNA hybrid, and D. Botstein for radioactively labeled P22 phage. We also thank J. M. Bishop, M. Green, and S. Spiegelman for manuscripts prior to publication. This work was supported by Public Health Service grant AI-08388 from the National Institute of Allergy and Infectious Diseases and by grant E-512 from the American Cancer Society. K. M. was a post-doctoral fellow of and D. B. received a faculty research award from the American Cancer Society.|
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|Deposited On:||19 May 2006|
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