Lukashevich I S, Lemeshko N N, Stel'makh T A, Golubev V P
Vopr Virusol. 1987 Jul-Aug;32(4):473-80.
Lassa virus purified in the isodensity sucrose concentration gradient had the following buoyant densities: 1.17 g/cm3 (sucrose), 1.19 g/cm3 (cesium chloride), 1.16 g/cm3 (urografin). Similar parameters were obtained for Machupo virus. Virion RNAs of these viruses contained 5 sedimentation classes of molecules: 30-31S, 28S, 22-24S, 18S, and 4-6S. Experiments on hybridization of individual sedimentation classes of RNA with an excess of poly(A)-containing RNA from the infected cells as well as inhibition of synthesis of 28S and 18S virion RNAs with low concentrations (0.005-0.5 micrograms/ml) of actinomycin D showed the genetic information for virus proteins to be coded for in two segments: 30-31S and 22-24S. The method of self-annealing demonstrated molecules with complementary sequence ("plus" and "minus" strands) in genome RNAs. In addition to previously described major proteins (78K, 64K, 37K), high performance liquid gel-penetrating chromatography of Machupo virus structural proteins revealed a minor protein with molecular weight of 50 kilodaltons. Pulse-chase experiments demonstrated in the infected cells a precursor-product metabolic bond between glycosylated proteins 78K and 37K. Lassa virus contained 3 structural major proteins with molecular weights 60, 48, and 34 kilodaltons (K). The 60K protein was detected in the nucleocapsid fraction, and 48 K protein in the soluble subvirion fraction. Proteins 60K and 34K were immunoprecipitated in greatest amounts in the infected cells.
1.17 g/cm³(蔗糖)、1.19 g/cm³(氯化铯)、1.16 g/cm³(泛影葡胺)。马丘波病毒也获得了类似参数。这些病毒的病毒粒子RNA包含5种沉降类别的分子:30 - 31S、28S、22 - 24S、18S和4 - 6S。用来自感染细胞的过量含聚(A)RNA对RNA的各个沉降类别进行杂交实验,以及用低浓度(0.005 - 0.5微克/毫升)放线菌素D抑制28S和18S病毒粒子RNA的合成,结果表明病毒蛋白的遗传信息由两个片段编码:30 - 31S和22 - 24S。自退火方法证明基因组RNA中存在具有互补序列的分子(“正”链和“负”链)。除了先前描述的主要蛋白质(78K、64K、37K)外,马丘波病毒结构蛋白质的高效液相凝胶渗透色谱显示出一种分子量为50千道尔顿的次要蛋白质。脉冲追踪实验在感染细胞中证明了糖基化蛋白78K和37K之间的前体 - 产物代谢关系。拉沙病毒包含3种分子量分别为60、48和34千道尔顿(K)的主要结构蛋白。60K蛋白在核衣壳部分被检测到,48K蛋白在可溶性亚病毒粒子部分被检测到。60K和34K蛋白在感染细胞中的免疫沉淀量最大。