Mrkic B, Kempf C
Institute of Biochemistry, University of Bern, Switzerland.
Arch Virol. 1996;141(10):1805-21. doi: 10.1007/BF01718196.
The fate of Semliki Forest virus (SFV) nucleocapsid, especially the capsid protein (C-protein), was investigated during the early stages of a productive infection in mosquito Aedes albopictus cells. Infection of the cells resulted in a time dependent accumulation of a C-protein derived fragment. This fragmentation of incoming viral nucleocapsid was prevented by NH4Cl, an agent generally used to elevate the pH in acidic intracellular compartments, suggesting that a low intravesicular pH is required for this process. Density gradient analysis of the postnuclear cell lysate demonstrated that the fragmentation was associated with a cellular compartment showing a density of 1.14 +/- 0.02 g/ml. This cellular compartment was devoid from a lysosomal marker enzyme and represented the timely preceding cellular fraction through which SFV passed before encountering a lysosomal fraction. Furthermore, the intracellular distribution of the viral, 3H-uridine-labeled RNA suggested that the same fraction might represent a key cellular compartment in which the separation of the viral RNA from the viral structural proteins is primed. In conclusion, these data lead to the suggestion that the fragmentation of incoming SFV nucleocapsids in Aedes albopictus cells might be the part of the mechanism leading to the release of viral RNA into the cytosol during early stages of productive infection.
在白纹伊蚊细胞进行有效感染的早期阶段,研究了塞姆利基森林病毒(SFV)核衣壳的命运,特别是衣壳蛋白(C蛋白)的命运。细胞感染导致C蛋白衍生片段随时间积累。NH4Cl可阻止进入细胞的病毒核衣壳的这种碎片化,NH4Cl是一种通常用于提高酸性细胞内区室pH值的试剂,这表明该过程需要低泡内pH值。对核后细胞裂解物的密度梯度分析表明,碎片化与密度为1.14±0.02 g/ml的细胞区室有关。该细胞区室缺乏溶酶体标记酶,代表了SFV在遇到溶酶体部分之前经过的及时的前一个细胞部分。此外,病毒的3H-尿苷标记RNA的细胞内分布表明,同一部分可能代表一个关键的细胞区室,在该区域中病毒RNA与病毒结构蛋白的分离开始启动。总之,这些数据表明,白纹伊蚊细胞中进入的SFV核衣壳的碎片化可能是在有效感染早期导致病毒RNA释放到细胞质中的机制的一部分。