Abstract
Two structurally distinct forms of RNA-directed DNA polymerase from avian myeloblastosis virus were resolved by chromatography on phosphocellulose and purified. In addition to RNA-directed DNA polymerase activity, both enzymes had ribonuclease H (RNase H) activity, which degraded the RNA moiety of RNA·DNA hybrids. As determined by sodium dodecyl sulfate-polyacrylamide disc-gel electrophoresis, one form had two subunits, alpha (α) and beta (β), with molecular weights of 65,000 and 105,000, respectively. The other had a single subunit, α, with a molecular weight of 65,000. The sedimentation coefficients of αβ and α, determined by glycerol gradient centrifugation in 0.35 M KCl, were 7.8 S and 5.2 S, respectively. Both enzymes had similar antigenic determinants and could not be distinguished by a differential response to several different RNA and DNA templates. We suggest that α, which contains both RNA-directed DNA polymerase and RNase H activity, is derived by dissociation of αβ; the function of the β subunit is unknown.
Keywords: RNA virus, polyacrylamide gel electrophoresis
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