Nugent C I, Kirkegaard K
Department of Molecular, Cellular and Developmental Biology, Howard Hughes Medical Institute, University of Colorado, Boulder 80309.
J Virol. 1995 Jan;69(1):13-22. doi: 10.1128/JVI.69.1.13-22.1995.
The mechanism of encapsidation of the RNA genome of poliovirus and other picornaviruses is unknown. To test whether any of the putative assembly intermediates of poliovirus could interact directly with the poliovirus RNA genome, poliovirus RNA was attached to magnetic streptavidin beads and incubated with partially purified extracts containing 35S-labeled 14S pentamer and 75S empty-capsid subviral particles from infected cells. The amount of labeled protein bound to the beads was monitored, thus testing the RNA-binding activities of only the labeled viral proteins in the preparations. In this assay, nonspecific RNA-binding activity was displayed by the 14S pentameric particles and mature virons. 75S empty capsids displayed no propensity to associate with RNA. 14S pentamers were demonstrated to form rapidly sedimenting complexes and to undergo a conformational alteration upon RNA binding. These findings are consistent with a direct role for the 14S pentameric particles in RNA packaging during poliovirus morphogenesis.
脊髓灰质炎病毒及其他小核糖核酸病毒的RNA基因组衣壳化机制尚不清楚。为了测试脊髓灰质炎病毒的任何假定组装中间体是否能直接与脊髓灰质炎病毒RNA基因组相互作用,将脊髓灰质炎病毒RNA连接到磁性链霉亲和素珠上,并与含有来自感染细胞的35S标记的14S五聚体和75S空衣壳亚病毒颗粒的部分纯化提取物一起孵育。监测与珠子结合的标记蛋白的量,从而仅测试制剂中标记病毒蛋白的RNA结合活性。在该试验中,14S五聚体颗粒和成熟病毒体表现出非特异性RNA结合活性。75S空衣壳没有显示出与RNA结合的倾向。已证明14S五聚体形成快速沉降的复合物,并在RNA结合时发生构象改变。这些发现与14S五聚体颗粒在脊髓灰质炎病毒形态发生过程中RNA包装中的直接作用一致。