Schwartz L B, Lawrence C, Thach R E, Roeder R G
J Virol. 1974 Sep;14(3):611-9. doi: 10.1128/JVI.14.3.611-619.1974.
The effect of encephalomyocarditis virus infection of MOPC 460 mouse plasmacytoma cells on host RNA synthesis and RNA polymerases was investigated. Consistent with work performed in other virus host systems, rates of RNA synthesis appeared to be inhibited in infected cells, whereas RNA degradation appeared normal. These results were further extended with isolated nuclei, in which distinct RNA polymerase activities could be studied under conditions where problems with RNA turnover and endogenous nucleotide pool sizes were insignificant. Endogenous nuclear RNA polymerase II activity was inhibited early postinfection and at 1 to 2 h prior to endogenous RNA polymerase I plus III activity. However, the solubilized enzymes were fully active with exogenous DNA as template. In fact, the levels of RNA polymerases I, II, and III, isolated from infected cells and nuclei, were indistinguishable from levels in uninfected cells and nuclei at each stage of their partial purification procedure. The chromatographic properties of the enzymes on DEAE-Sephadex were also unaltered. Furthermore, the RNA synthetic activity of these isolated enyzmes, or of nuclei isolated from uninfected cells, was resistant to extracts of nuclei or of cytoplasmic fractions from infected cells. These results are discussed in terms of a possible inhibition of RNA synthesis in vivo at the level of transcription initiation.
研究了脑心肌炎病毒感染MOPC 460小鼠浆细胞瘤细胞对宿主RNA合成及RNA聚合酶的影响。与在其他病毒宿主系统中开展的工作一致,受感染细胞中的RNA合成速率似乎受到抑制,而RNA降解看起来正常。利用分离的细胞核进一步拓展了这些结果,在这些细胞核中,可以在RNA周转和内源性核苷酸池大小问题不显著的条件下研究不同的RNA聚合酶活性。内源性核RNA聚合酶II活性在感染后早期以及在内源性RNA聚合酶I加III活性出现之前1至2小时受到抑制。然而,溶解的酶以外源DNA为模板时具有完全活性。实际上,在部分纯化过程的每个阶段,从受感染细胞和细胞核中分离出的RNA聚合酶I、II和III的水平与未感染细胞和细胞核中的水平没有区别。这些酶在DEAE-葡聚糖上的色谱特性也未改变。此外,这些分离酶或从未感染细胞中分离出的细胞核的RNA合成活性对来自受感染细胞核提取物或细胞质部分具有抗性。从转录起始水平对体内RNA合成可能受到的抑制方面对这些结果进行了讨论。