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首页> 外文期刊>Journal of Virology >Aphidicolin inhibition of the production of replicative-form DNA during bovine parvovirus infection.
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Aphidicolin inhibition of the production of replicative-form DNA during bovine parvovirus infection.

机译：蚜虫蛋白抑制在牛节目病毒感染期间的复制形式DNA的产生。

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Since parvoviruses apparently do not possess a DNA polymerase activity, one or more of the host cell DNA polymerases must be responsible for replicating the single-stranded DNA genome. We have focused on determining which polymerase, alpha, beta, or gamma (pol alpha, pol beta, or pol gamma, respectively), is responsible for the first step in bovine parvoviral DNA replication: conversion of the single-stranded DNA genome to a parental replicative form (RF). In this study, we used aphidicolin, a specific inhibitor of DNA pol alpha, to assay for the requirement of pol alpha activity in parental RF formation in vivo. Synchronized cell cultures were infected with bovine parvovirus with or without aphidicolin, and the products of viral replication were separated on agarose gels and identified by Southern blot analysis. We found that complete inhibition of viral DNA synthesis resulted when 20 microM aphidicolin was present throughout the infection. In addition, viral DNA synthesis was inhibited by as little as 1 microM aphidicolin, whereas lower concentrations (0.1 and 0.01 microM) resulted in partial inhibition of the replication process. Using 32P-labeled bovine parvovirus as the input virus we differentiated parental RF from daughter RF and progeny DNA synthesis. We conclude that DNA pol alpha is required for the production of RF during bovine parvovirus replication in vivo and that this requirement is most likely for the conversion of bovine parvovirus input single-stranded DNA to parental RF. These results do not rule out a possible role for DNA pol gamma in the first step, nor do they rule out a role for pol alpha or pol gamma in later stages of the replication cycle.
机译：由于虚条病毒显然不具有DNA聚合酶活性，因此宿主细胞DNA聚合酶中的一个或多个必须是转换单链DNA基因组的原因。我们专注于确定哪种聚合酶，α，β或γ（Polα，Polβ或Polγ），其负责牛节目病毒DNA复制的第一步：将单链DNA基因组的转化为a父母复制形式（RF）。在该研究中，我们使用蚜虫蛋白，DNA Polα的特异性抑制剂，用于测定体内亲本RF形成中POLα活性的要求。同步细胞培养物用或没有蚜虫蛋白的牛剖视病毒感染，并且在琼脂糖凝胶上分离病毒复制产物并通过Southern印迹分析鉴定。我们发现当在整个感染过程中存在20微米蚜虫蛋白时，对病毒性DNA合成的完全抑制产生。此外，抑制病毒DNA合成只要少至1微米蚜虫蛋白，而较低浓度（0.1和0.01微米）导致部分抑制复制过程。使用32P标记的牛Parvovirus作为输入病毒，我们将父母RF与女儿RF和后代DNA合成分化。我们得出结论，在体内生产RF期间RF需要DNA POLα，并且该要求最有可能转化牛节目病毒输入单链DNA至亲本RF。这些结果不会在第一步中排除DNAPolγ的可能作用，也不会在复制周期的后期阶段排列Pol alpha或Pol伽马的作用。

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