Hirsch I, Vonka V
J Virol. 1974 Jun;13(6):1162-8. doi: 10.1128/JVI.13.6.1162-1168.1974.
Cells of a continuous cell line derived from rabbit embryo fibroblasts were infected with herpes simplex type 1 virus (HSV-1) and maintained in the presence of either [5-(3)H]uridine or [methyl-(3)H]thymidine or (32)PO(4) (3-). Nucleocapsids were isolated from the cytoplasmic fraction, partially purified, and treated with DNase and RNase. From the pelleted nucleocapsids, DNA was extracted and purified by centrifugation in sucrose and cesium sulfate gradients. The acid-precipitable radioactivity of [5-(3)H]uridine-labeled DNA was partially susceptible to pancreatic RNase and alkaline treatment; the susceptibility to the enzyme decreased with increasing salt concentration. No drop of activity of DNA labeled with [(3)H]thymidine was observed either after RNase or alkali treatment. Base composition analysis of [5-(3)H]uridine-labeled DNA showed that the radioactivity was recovered as uracil and cytosine. In the cesium sulfate gradient, the purified [5-(3)H]uridine-labeled DNA banded at the same position as the (32)P-labeled DNA. The present data tend to suggest that ribonucleotide sequences are present in HSV DNA, that they are covalently attached to the viral DNA, and that they can form double-stranded structures.
源自兔胚胎成纤维细胞的连续细胞系的细胞被单纯疱疹病毒1型（HSV-1）感染，并在[5-(3)H]尿苷或[甲基-(3)H]胸苷或(32)PO(4) (3-)存在的情况下进行培养。从细胞质部分分离出核衣壳，进行部分纯化，并用DNA酶和RNA酶处理。从沉淀的核衣壳中提取DNA，并通过在蔗糖和硫酸铯梯度中离心进行纯化。[5-(3)H]尿苷标记的DNA的酸沉淀放射性对胰RNA酶和碱性处理部分敏感；随着盐浓度的增加，对该酶的敏感性降低。在用RNA酶或碱处理后，未观察到[(3)H]胸苷标记的DNA活性下降。[5-(3)H]尿苷标记的DNA的碱基组成分析表明，放射性以尿嘧啶和胞嘧啶的形式回收。在硫酸铯梯度中，纯化的[5-(3)H]尿苷标记的DNA与(32)P标记的DNA在相同位置形成条带。目前的数据倾向于表明，核糖核苷酸序列存在于HSV DNA中，它们与病毒DNA共价连接，并且它们可以形成双链结构。
