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首页> 外文期刊>Journal of Virology >Synthesis and Intracellular Localization of Vaccinia Virus Deoxyribonucleic Acid-dependent Ribonucleic Acid Polymerase
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Synthesis and Intracellular Localization of Vaccinia Virus Deoxyribonucleic Acid-dependent Ribonucleic Acid Polymerase

机译：痘苗病毒脱氧核糖核酸依赖性核糖核酸聚合酶的合成与细胞内定位

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摘要

The time course of vaccinia deoxyribonucleic acid (DNA)-dependent ribonucleic acid (RNA) polymerase synthesis and its intracellular localization were studied with virus-infected HeLa cells. Viral RNA polymerase activity could be meassured shortly after viral infection in the cytoplasmic fraction of infected cells in vitro. However, unless the cells were broken in the presence of the nonionic detergent Triton-X-100, no significant synthesis of new RNA polymerase was detected during the viral growth cycle. When cells were broken in the presence of this detergent, extensive increases in viral RNA polymerase activity were observed late in the infection cycle. The onset of new RNA polymerase synthesis was dependent on prior viral DNA replication. Fluorodeoxyuridine (5 × 10?5m) prevented the onset of viral polymerase synthesis. Streptovitacin A, a specific and complete inhibitor of protein synthesis in HeLa cells, prevented the synthesis of RNA polymerase. Thus, the synthesis of RNA polymerase is a “late” function of the virus. The newly synthesized RNA polymerase activity was primarily bound to particles which sedimented during high-speed centrifugation. These particles have been characterized by sucrose gradient centrifugation. A major class of active RNA polymerase particles were considerably “lighter” than whole virus in sucrose gradients. These particles were entirely resistant to the action of added pancreatic deoxyribonuclease, and they were not stimulated by added calf thymus primer DNA. It is concluded that these particles are not active in RNA synthesis in vivo, and that activation occurs as a result of detergent treatment in vitro.
机译：用病毒感染的HeLa细胞研究了疫苗脱氧核糖核酸（DNA）依赖性核糖核酸（RNA）聚合酶合成及其细胞内定位的时间过程。在体外感染细胞的细胞质级分中病毒感染后不久可能测量病毒RNA聚合酶活性。然而，除非在存在非离子洗涤剂Triton-X-100存在下细胞被破坏，否则在病毒生长循环期间未检测到新的RNA聚合酶的显着合成。当在这种洗涤剂存在下细胞被破坏时，在感染循环中晚期观察到病毒RNA聚合酶活性的广泛增加。新RNA聚合酶合成的发作取决于现有的病毒DNA复制。氟脱氧尿苷（5×10 β5 m）防止了病毒聚合酶合成的发作。 Streptovitacin A，HeLa细胞中蛋白质合成的特异性和完整抑制剂，防止了RNA聚合酶的合成。因此，RNA聚合酶的合成是病毒的“晚期”功能。新合成的RNA聚合酶活性主要与在高速离心过程中沉积的颗粒结合。这些颗粒的特征在于蔗糖梯度离心。主要类活性RNA聚合酶颗粒比蔗糖梯度的整个病毒相当于“更轻”。这些颗粒完全抵抗加入的胰脱氧氧核酸酶的作用，并且不会通过添加小牛胸腺引物DNA刺激它们。结论是，这些颗粒在体内的RNA合成中不活跃，并且在体外洗涤剂处理的结果发生活化。

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