Ooka T, Lenoir G, Daillie J
J Virol. 1979 Jan;29(1):1-10. doi: 10.1128/JVI.29.1.1-10.1979.
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 alpha- and beta-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 polymerases 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 properteis similar to those reported for other herpesvirus DNA polymerases. The enzyme exhibited optimal activity for copying activated calf DNA in the presence of 50 mH (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)10 and oligo(dT)10, indicating that this enzyme has characters distinct from DNA polymerase gamma, reverse transcriptase, and terminal deoxynucleotidyl transferase. Phosphonacetic acid inhibited not only EBV DNA polymerase, but also, to a lesser degree, the cellular polymerase alpha. AraATP did not severely inhibit viral activity, whereas the polymerase alpha was inhibited most effectively. Both EBV polymerase and polymerase alpha were inhibited at a comparable level by araCTP.
在P3HR-I细胞培养物中添加碘脱氧尿苷，导致爱泼斯坦-巴尔病毒（EBV）诱导的DNA聚合酶活性和早期抗原阳性细胞大幅增加。通过在琼脂糖6B、二乙氨基乙基纤维素和磷酸纤维素上进行连续柱层析，将这种EBV诱导的DNA聚合酶与细胞α-和β-聚合酶分离，得到约320倍的部分纯化。部分纯化的EBV DNA聚合酶可通过其对盐的激活作用、催化特性以及对N-乙基马来酰亚胺、膦甲酸、araATP和araCTP的敏感程度与细胞DNA聚合酶区分开来。该病毒聚合酶表现出与其他疱疹病毒DNA聚合酶报道的特性相似。该酶在存在50 mM硫酸铵时对复制活化的小牛DNA表现出最佳活性，并且对150 mM硫酸铵具有抗性。它能高效利用模板引物聚（dC）-寡聚（dG）12 - 18或聚（dA）-寡聚（dT）12 - 18，但无法复制聚（rA）-寡聚（dT）10和寡聚（dT）10，这表明该酶具有与DNA聚合酶γ、逆转录酶和末端脱氧核苷酸转移酶不同的特性。膦甲酸不仅抑制EBV DNA聚合酶，还在较小程度上抑制细胞聚合酶α。AraATP不会严重抑制病毒活性，而聚合酶α受到最有效的抑制。EBV聚合酶和聚合酶α都受到araCTP相当程度的抑制。
