Hirai K, Watanabe Y
Biochim Biophys Acta. 1976 Oct 18;447(3):328-39. doi: 10.1016/0005-2787(76)90056-3.
Productive infection of WI-38 cells with human cytomegalovirus (HCMV) induced the increase in the activity of DNA polymerases as well as the synthesis of viral and cellular DNA. Sedimentation analyses in sucrose gradients of high ionic strength showed that the HCMV infection caused marked increase in the activity of alpha-type polymerases (resolved into alpha1, 8 S, and alpha 2, 6 S, in the present experiments), while the infection little affected the level of beta-type polymerase (about 3.5 S) activity in both the nuclei and cytoplasm. Such increase in alpha-type polymerases was also observed when DNA synthesis in WI-38 cells was enhanced by SV40 infection or by an increased concentration of serum in medium. Phosphonacetate, which selectively blocked the synthesis of HCMV DNA, did not significantly affect the HCMV-mediated induction of DNA polymerases. However, phosphonoacetate added in the reaction mixture for DNA polymerase assay inhibited the activity of the HCMV-induced polyperase alpha, but not of the polymerases alpha2 and beta. These results support the idea that alpha-type polymerases are involved in the replicative synthesis of cellular and viral DNA.
人巨细胞病毒(HCMV)对WI-38细胞的有效感染导致DNA聚合酶活性增加以及病毒和细胞DNA的合成。在高离子强度的蔗糖梯度中进行沉降分析表明,HCMV感染导致α型聚合酶(在本实验中解析为α1, 8 S和α2, 6 S)的活性显著增加,而感染对细胞核和细胞质中β型聚合酶(约3.5 S)的活性水平影响很小。当通过SV40感染或培养基中血清浓度增加来增强WI-38细胞中的DNA合成时,也观察到α型聚合酶的这种增加。选择性阻断HCMV DNA合成的膦乙酸对HCMV介导的DNA聚合酶诱导没有显著影响。然而,在用于DNA聚合酶测定的反应混合物中添加膦乙酸会抑制HCMV诱导的聚合酶α的活性,但不会抑制聚合酶α2和β的活性。这些结果支持α型聚合酶参与细胞和病毒DNA复制合成的观点。