Zhirnov O P, Bukrinskaia A G
Vopr Virusol. 1981 Nov-Dec(6):667-77.
Subcellular distribution and intracellular transportation of influenza virus (WSN) proteins were studied in chick embryo cell culture. After cumulative labeling with 14C-amino acids proteins M, NSI, NP, P1, P2, P3 were found in the nuclei. In addition to these proteins, the cytoplasm contained viral hemagglutinin (HA). The content of P1, P3 and particularly P2 proteins as compared with NP was much lower in the nuclei than in the cytoplasm. P1 and P3 proteins recovered from the nuclei had a higher electrophoretic mobility in gel than similar proteins from the cytoplasmic RNP and from the virus. HA, M, and NSI proteins in the cytoplasm were detected mainly in the membrane fraction, and NP, P1, P2, and P3 in viral RNP some of which were associated with the membranes. Analysis of proteins in polyacrylamide gel showed protein P3 in RNPs associated with membranes to have a higher mobility than that in free viral RNPs. Protein P2 also accumulated in significant amounts in the cytoplasm in the soluble form. The intracellular transport of proteins was studied by the pulse-chase technique at two intervals: 3 1/2 and 6 1/2 hours postinfection. NP, M, NSI, P1, P2, and P3 proteins migrated into the nuclei at a different relative rate. The rate of migration into the nuclei of individual proteins (particularly P1, P2, and M) changed in the course of infection. A large portion of molecules of P1 and P3 proteins was present in the nuclei for 1 1/2 - 2 hours after synthesis only. The newly synthesized NP, P1, P2, and P3 proteins integrated into the free cytoplasmic RNP of the virus asynchronously.
在鸡胚细胞培养中研究了流感病毒(WSN)蛋白的亚细胞分布和细胞内运输。用¹⁴C -氨基酸进行累积标记后,在细胞核中发现了M、NSI、NP、P1、P2、P3蛋白。除了这些蛋白外,细胞质中含有病毒血凝素(HA)。与NP相比,细胞核中P1、P3尤其是P2蛋白的含量远低于细胞质。从细胞核中回收的P1和P3蛋白在凝胶中的电泳迁移率高于来自细胞质核糖核蛋白(RNP)和病毒的类似蛋白。细胞质中的HA、M和NSI蛋白主要在膜部分被检测到,而NP、P1、P2和P3在病毒RNP中,其中一些与膜相关。聚丙烯酰胺凝胶中的蛋白分析表明,与膜相关的RNP中的P3蛋白迁移率高于游离病毒RNP中的P3蛋白。P2蛋白也以大量可溶形式在细胞质中积累。通过脉冲追踪技术在感染后的两个时间间隔(3.5小时和6.5小时)研究了蛋白的细胞内运输。NP、M、NSI、P1、P2和P3蛋白以不同的相对速率迁移到细胞核中。在感染过程中,单个蛋白(特别是P1、P2和M)迁移到细胞核的速率发生了变化。P1和P3蛋白的大部分分子仅在合成后1.5 - 2小时存在于细胞核中。新合成的NP、P1、P2和P3蛋白异步整合到病毒的游离细胞质RNP中。