Hackett P B, Egberts E, Traub P
Eur J Biochem. 1978 Feb;83(2):341-52. doi: 10.1111/j.1432-1033.1978.tb12100.x.
We have prepared homologous, fractionated, cell-free translational systems from uninfected and mengovirus-infected Ehrlich ascites tumor cells in order to determine what alterations occur following virus infection in the translational machinery of the host cell. Two major differences distinguish the system developed from infected cells. First, it has a 40% lower rate of protein synthesis, primarily a consequence of the rate of chain elongation, which is depressed to 60 amino acids/min from 90 amino acids/min in the system from uninfected cells. Second, at supraoptimal concentrations of Mg2+ and K+ the system from virus-infected cells supports the translation of mengovirus RNA but not host mRNA. These differences between the two systems may reflect specific changes which are responsible for the selective translation of mengovirus RNA in the infected cell. In both systems the optimal concentrations of polyamines, monovalent and divalent cations, mRNA, and ribosomal subunits are the same for the translation of either host or viral RNA. This uniformity is useful in experiments, designed to investigate the selective translation of viral RNA, where various components of the two systems are interchanged.
我们从未感染和感染脑心肌炎病毒的艾氏腹水瘤细胞中制备了同源、分级分离的无细胞翻译系统,以确定宿主细胞翻译机制在病毒感染后会发生哪些变化。从感染细胞中开发的系统有两个主要区别。首先,其蛋白质合成速率降低了40%,这主要是链延伸速率降低的结果,在未感染细胞的系统中,链延伸速率从每分钟90个氨基酸降至每分钟60个氨基酸。其次,在Mg2+和K+的超最佳浓度下,来自病毒感染细胞的系统支持脑心肌炎病毒RNA的翻译,但不支持宿主mRNA的翻译。这两个系统之间的这些差异可能反映了导致感染细胞中脑心肌炎病毒RNA选择性翻译的特定变化。在两个系统中,多胺、单价和二价阳离子、mRNA和核糖体亚基的最佳浓度对于宿主或病毒RNA的翻译都是相同的。这种一致性在旨在研究病毒RNA选择性翻译的实验中很有用,在这些实验中,两个系统的各种成分会相互交换。