Nishioka Y, Silverstein S
J Virol. 1978 Sep;27(3):619-27. doi: 10.1128/JVI.27.3.619-627.1978.
We describe experiments which demonstrate that shortly after infection of Friend erythroleukemia cells with herpes simplex virus (HSV), polyribosomes dissociate and cellular mRNA degrades. Analysis of infected cell extracts on sucrose density gradients demonstrates that the majority of the polyribosomes have dissociated to monoribosomes at 2 h postinfection. Physical measurements of infected-cell RNAs support this conclusion and demonstrate that the polyadenylated RNAs decrease in size. The degradation of mRNA is apparently a stochastic process as judged by the failure to detect a shift in the Crt1/2 when polyadenylated RNA extracted from infected cells at different times is hybridized to globin complementary DNA. In experiments designed to determine whether dissociation of polyribosomes is sufficient to cause degradation of globin mRNA, the amount of globin mRNA in uninfected cells did not change when cells were treated with NaF or pactamycin at concentrations sufficient to dissociate all polyribosomes. In cells infected with UV-irradiated virus polyribosomes dissociate but globin mRNA does not degrade, suggesting that it is possible to separate dissociation from degradation.
我们描述了一些实验,这些实验表明,在用单纯疱疹病毒(HSV)感染弗瑞德红白血病细胞后不久,多核糖体就会解离,细胞mRNA也会降解。在蔗糖密度梯度上对感染细胞提取物进行分析表明,在感染后2小时,大多数多核糖体已解离为单核糖体。对感染细胞RNA的物理测量支持了这一结论,并表明多聚腺苷酸化RNA的大小减小。从不同时间提取的感染细胞的多聚腺苷酸化RNA与珠蛋白互补DNA杂交时,未能检测到Crt1/2的变化,由此判断mRNA的降解显然是一个随机过程。在旨在确定多核糖体的解离是否足以导致珠蛋白mRNA降解的实验中,当用足以解离所有多核糖体的浓度的氟化钠或放线菌酮处理未感染细胞时,未感染细胞中珠蛋白mRNA的量没有变化。在用紫外线照射的病毒感染的细胞中,多核糖体解离,但珠蛋白mRNA不降解,这表明有可能将解离与降解分开。