Arlinghaus R B, Polatnick J
Proc Natl Acad Sci U S A. 1969 Mar;62(3):821-8. doi: 10.1073/pnas.62.3.821.
The foot-and-mouth disease virus-RNA polymerase complex was released from membrane particulates present in the cytoplasm of infected baby hamster kidney cells. The soluble polymerase complex was fractionated by zonal centrifugation in sucrose gradients. Two polymerase complexes (RNA and protein complex) active in the cell-free system were isolated and had S-rate ranges of 20-70S and 100-300S, respectively. The light polymerase complex contained 20S double-stranded RNA; and the heavy polymerase complex contained a polydisperse, partially RNase-resistant RNA. The cell-free product of these two polymerase complexes was analyzed by zonal centrifugation in sucrose gradients. The light polymerase complex synthesized only 20S double-stranded RNA. The product of the heavy polymerase complex contained no detectable 20S double-stranded RNA and only a peak of single-stranded RNA with S-rate corresponding to 37S viral RNA. A third polymerase complex was isolated with S-rate greater than 300S, and it contained a polydisperse, partially RNase-resistant RNA. This third polymerase complex synthesized both 37S viral RNA and 20S double-stranded RNA in the cell-free system, and it is probably the native polymerase complex still bound to cellular particulates.
口蹄疫病毒-RNA聚合酶复合物是从感染的幼仓鼠肾细胞胞质中的膜颗粒中释放出来的。可溶性聚合酶复合物通过蔗糖梯度区带离心进行分级分离。分离出了在无细胞系统中具有活性的两种聚合酶复合物(RNA和蛋白质复合物),其沉降速率范围分别为20-70S和100-300S。较轻的聚合酶复合物含有20S双链RNA;较重的聚合酶复合物含有一种多分散的、部分抗核糖核酸酶的RNA。通过蔗糖梯度区带离心对这两种聚合酶复合物的无细胞产物进行分析。较轻的聚合酶复合物仅合成20S双链RNA。较重的聚合酶复合物的产物未检测到20S双链RNA,只有一个沉降速率对应于37S病毒RNA的单链RNA峰。分离出了第三种沉降速率大于300S的聚合酶复合物,它含有一种多分散的、部分抗核糖核酸酶的RNA。这种第三种聚合酶复合物在无细胞系统中既合成37S病毒RNA也合成20S双链RNA,它可能是仍与细胞颗粒结合的天然聚合酶复合物。