Bossart W, Egger D, Rasser Y, Bienz K
Intervirology. 1984;21(3):150-8. doi: 10.1159/000149513.
Poliovirus (type 1, Mahoney) proteins preferentially inhibited in vitro RNA polymerase II activity in uninfected isolated HEp-2 cell nuclei, as demonstrated by electron microscopic autoradiography. Structural integrity of the nuclei and preincubation of nuclei and poliovirus proteins in vitro prior to addition of [3H]-UTP were prerequisites for differential inhibition. During in vitro incubation, [3H]-labeled poliovirus proteins were shown to accumulate in the uninfected isolated nuclei. Similar poliovirus proteins accumulated in the nuclei to higher relative amounts as compared to the outside or were excluded from the nuclei after in vitro incubation, as in intact poliovirus-infected cells. Since the in vitro transcription system of isolated nuclei exhibited two characteristics of poliovirus infection, i.e., preferential inhibition of RNA polymerase II activity and accumulation of poliovirus proteins in the nuclei, it may be useful to study host RNA synthesis inhibition.
如电子显微镜放射自显影所示,脊髓灰质炎病毒(1型,马奥尼株)蛋白在体外优先抑制未感染的分离的HEp-2细胞核中的RNA聚合酶II活性。细胞核的结构完整性以及在添加[3H]-UTP之前细胞核与脊髓灰质炎病毒蛋白在体外的预孵育是差异抑制的先决条件。在体外孵育期间,[3H]标记的脊髓灰质炎病毒蛋白显示在未感染的分离细胞核中积累。与外部相比,类似的脊髓灰质炎病毒蛋白在细胞核中积累到更高的相对量,或者在体外孵育后被排除在细胞核之外,如同在完整的脊髓灰质炎病毒感染细胞中一样。由于分离细胞核的体外转录系统表现出脊髓灰质炎病毒感染的两个特征,即RNA聚合酶II活性的优先抑制和脊髓灰质炎病毒蛋白在细胞核中的积累,因此它可能有助于研究宿主RNA合成抑制。