In vitro polyoma DNA synthesis: Involvement of RNA in discontinuous chain growth |
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Authors: | Tony Hunter Bertold Francke |
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Affiliation: | The Salk Institute for Biological Studies Post Office Box 1809 San Diego, Calif. 92112 U.S.A. |
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Abstract: | Short fragments of DNA (5 S) isolated by denaturation from polyoma replicative intermediates pulse-labeled in vitro were shown to have RNA covalently attached by three criteria: (1) such fragments were slightly denser than bulk viral DNA. (2) They could be labeled directly with α-32P-labeled ribotriphosphates. (3) Alkaline hydrolysis of fragments labeled with α-32P-labeled deoxynucleoside triphosphates showed 32P transfer to 3′ ribonucleoside monophosphates. Except for a preference of transfer from dC, the link showed little sequence specificity. The data are compatible with the notion that all short fragments in replicating viral DNA are initiated by an RNA primer. This RNA is maximally 30 bases long and is rather short-lived. |
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