Krishnan I, Baglioni C
Mol Cell Biol. 1981 Oct;1(10):932-8. doi: 10.1128/mcb.1.10.932-938.1981.
(2'-5')Oligoadenylic acid [(2'-5')An] polymerase activity was measured in extracts of human lymphoblastoid cells of the Namalva line cultured under different conditions. Exponentially growing cells had a relatively low polymerase activity level, whereas cells grown to limit density showed elevated levels. When fresh medium was added to growth-arrested cells, (2'-5')An polymerase activity decreased concomitantly with the initiation of active deoxyribonucleic acid synthesis. An increase in polymerase activity level was also observed after exponentially growing cells were transferred from medium containing 20% serum to fresh medium containing 0.2% serum. These cells diminished deoxyribonucleic acid synthesis and remained quiescent until 20% serum was again added. Polymerase activity level decreased as the cells entered into S phase. The addition of the inhibitor of deoxyribonucleic acid synthesis, hydroxyurea, to exponentially growing cells did not increase polymerase level, indicating that cells blocked in S phase and at the G1-S boundary maintained the basal level of this enzyme. Degradation of labeled (2'-5')An was measured in extracts of Namalva cells cultured under different conditions, but no significant differences among degradative activities were observed. Since (2'-5')An polymerase activity is one of the enzymatic activities induced by interferon, we measured interferon titers in Namalva cell medium. Less than 1 reference unit per ml was detected in cells grown under different conditions. Moreover, the increase in (2'-5')An polymerase activity level in cells transferred from 20 to 0.2% serum was not prevented by including anti-lymphoblastoid interferon antibody in the medium. These results suggest that the activity level of (2'-5')An polymerase is regulated in Namalva cells on the basis of the growth status of the cells and that this regulatory mechanism is apparently not activated by interferon.
在不同条件下培养的Namalva系人淋巴母细胞提取物中,测定了(2'-5')寡腺苷酸[(2'-5')An]聚合酶活性。指数生长的细胞聚合酶活性水平相对较低,而生长至极限密度的细胞则显示出较高的水平。当向生长停滞的细胞中添加新鲜培养基时,(2'-5')An聚合酶活性随着活性脱氧核糖核酸合成的开始而相应降低。在指数生长的细胞从含20%血清的培养基转移至含0.2%血清的新鲜培养基后,也观察到聚合酶活性水平升高。这些细胞减少了脱氧核糖核酸合成并保持静止,直至再次添加20%血清。随着细胞进入S期,聚合酶活性水平降低。向指数生长的细胞中添加脱氧核糖核酸合成抑制剂羟基脲,并未提高聚合酶水平,这表明停滞在S期和G1-S边界的细胞维持了该酶的基础水平。在不同条件下培养的Namalva细胞提取物中,测定了标记的(2'-5')An的降解情况,但未观察到降解活性之间的显著差异。由于(2'-5')An聚合酶活性是干扰素诱导的酶活性之一,我们测定了Namalva细胞培养基中的干扰素滴度。在不同条件下生长的细胞中,每毫升检测到的干扰素滴度低于1个参考单位。此外,在培养基中加入抗淋巴母细胞干扰素抗体,并未阻止从20%血清转移至0.2%血清的细胞中(2'-5')An聚合酶活性水平的升高。这些结果表明,Namalva细胞中(2'-5')An聚合酶的活性水平根据细胞的生长状态进行调节,并且这种调节机制显然不受干扰素激活。