The addition of iododeoxyuridine to P3HR-I cell cultures led to a large increase in both Epstein-Barr virus (EBV)-induced DNA polymerase activity and early antigen-positive cells. This EBV-induced DNA polymerase was separated from the cellular a-and ,B-polymerases by sequential column chromatography on Sepharose 6B, DEAE-cellulose, and phosphocellulose, resulting in partial purification of about 320-fold. The partially purified-EBV DNA polymerase could be distinguished from the cellular DNA

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... merases by its activation by salts, its catalytic properties, and its degree of sensitivity to N-ethylmaleimide, phosphonoacetic acid, araATP, and araCTP. The viral polymerase showed properties similar to those reported for other herpesvirus DNA polymerases. The enzyme exhibited optimal activity for copying activated calf DNA in the presence of 50 mM (NH4)2SO4 and was resistant to 150 mM (NH4)2SO4. It utilized with high efficiency template-primer poly(dC)-oligo(dG)12 18 or poly(dA)-oligo(dT)12-18, but failed to copy poly(rA)-oligo(dT)1o and oligo(dT)IO, indicating that this enzyme has characters distinct from DNA polymerase y, reverse transcriptase, and terminal deoxynucleotidyl transferase. Phosphonoacetic acid inhibited not only EBV DNA polymerase, but also, to a lesser degree, the cellular polymerase a. AraATP did not severely inhibit viral activity, whereas the polymerase a was inhibited most effectively. Both EBV polymerase and polymerase a were inhibited at a comparable level by araCTP. rified 320-fold and the salt sensitivity, template specificity, and chemical inhibitor sensitivity were studied. on May 9, 2020 by guest