Moyer S A
Department of Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville 32610.
Virology. 1989 Sep;172(1):341-5. doi: 10.1016/0042-6822(89)90136-0.
We have tested the ability of heterologous viral proteins to support the in vivo and in vitro replication of the RNA of defective interfering (DI) particles of two serotypes of VSV and of Sendai virus. In all the combinations of heterologous coinfections in vivo, DI particle replication was observed only in the coinfection with the VSV-Indiana DI particle and wild-type VSV-New Jersey. By quantitating RNA synthesis in reconstitution experiments we showed that with DI nucleocapsids isolated from infected cells, however, the soluble protein fraction from heterologous wild-type virus-infected cells could substitute in vitro to varying degrees for the homologous proteins in the elongation reaction of RNA replication and encapsidation. In these cases successful replication was confirmed by demonstrating the specific association of the heterologous N protein with the product nucleocapsid RNA. The initiation step, that is, the initial binding of the nucleocapsid protein to the leader RNA, in contrast, requires the homologous protein, since heterologous viral proteins could not support RNA replication and encapsidation from purified DI particles.
我们测试了异源病毒蛋白支持两种血清型水疱性口炎病毒(VSV)和仙台病毒缺陷干扰(DI)颗粒RNA在体内和体外复制的能力。在体内所有异源共感染组合中,仅在VSV-印第安纳DI颗粒与野生型VSV-新泽西共感染时观察到DI颗粒复制。然而,通过在重组实验中对RNA合成进行定量,我们发现,从感染细胞中分离出DI核衣壳后,异源野生型病毒感染细胞的可溶性蛋白组分在体外可不同程度地替代同源蛋白参与RNA复制和衣壳化的延伸反应。在这些情况下,通过证明异源N蛋白与产物核衣壳RNA的特异性结合来确认成功复制。相比之下,起始步骤,即核衣壳蛋白与前导RNA的初始结合,需要同源蛋白,因为异源病毒蛋白无法支持从纯化的DI颗粒进行RNA复制和衣壳化。