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. 1974 Jul;14(1):86–98. doi: 10.1128/jvi.14.1.86-98.1974

Vaccinia Virus Polyriboadenylate Polymerase: Covalent Linkage of the Product with Polyribonucleotide and Polydeoxyribonucleotide Primers

Bernard Moss 1, Edith N Rosenblum 1
PMCID: PMC355481  PMID: 4836599

Abstract

A polyriboadenylate [poly(A)] polymerase, purified from vaccinia virus cores, was stimulated by polydeoxyriboadenylate: polydeoxyribothymidylate [poly(dA:dT)] and by polyribocytidylate [poly(C)] primers suggesting mechanisms of either transcription or terminal addition. Evidence for the latter was obtained by the demonstration of covalent linkages between the poly(A) products and both primers. In 99% dimethylsulfoxide-sucrose gradients, the sedimentation of poly(A) formed with poly(dA: dT) primer was reduced after DNase I treatment and the sedimentation of poly(A) formed with poly(C) primer was reduced by RNase A treatment, whereas the sedimentation of poly(A) formed without primer was not affected by either. Formation of a phosphodiester bond between primer and product was demonstrated by means of isotope transfer experiments. 32P from α-[32P]ATP was transferred to 2′(3′)-CMP after alkaline or enzymatic hydrolysis of the poly(C)-primed polymerase reaction product. Transfer primarily or exclusively to 3′-dTMP was found after enzymatic hydrolysis of the poly(dA: dT)-primed polymerase reaction product. The elution pattern of the poly(A) polymerase from DNA-cellulose suggested that a single enzyme catalyzes the attachment of adenylate residues to both polyribonucleotide and polydeoxyribonucleotide primers; nevertheless the purest enzyme preparations contain two bands resolved by polyacrylamide gel electrophoresis in sodium dodecyl sulfate.

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Selected References

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

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