Belov G A, Evstafieva A G, Rubtsov Y P, Mikitas O V, Vartapetian A B, Agol V I
M. P. Chumakov Institute of Poliomyelitis and Viral Encephalitides, Russian Academy of Medical Sciences, Moscow Region, Russia.
Virology. 2000 Sep 30;275(2):244-8. doi: 10.1006/viro.2000.0427.
A HeLa cell line expressing the green fluorescent protein fused to the SV40 T-antigen nuclear localization signal (EGFP-NLS) was established. Fluorescence in these cells was confined to the nuclei. After poliovirus infection, cytoplasmic fluorescence in a proportion of cells could be detected by 1 h postinfection (p.i.) and in virtually all of the fluorescent cells by 2 h p.i. The relocation could be prevented by cycloheximide but not by inhibition of poliovirus replication by guanidine. HCl. Nuclear exit of a protein composed of three copies of GFP fused to the NLS also occurred upon poliovirus infection. A similar redistribution of EGFP-NLS took place upon infection with coxsakievirus B3 and, to a lesser extent, with vesicular stomatitis virus. The EGFP-NLS efflux was not due to the loss of NLS. Thus, some positive-strand and negative-strand RNA viruses trigger a rapid nonspecific relocation of nuclear proteins.
建立了一种表达与SV40 T抗原核定位信号融合的绿色荧光蛋白(EGFP-NLS)的HeLa细胞系。这些细胞中的荧光局限于细胞核。脊髓灰质炎病毒感染后,感染后1小时(p.i.)可在一部分细胞中检测到细胞质荧光,到感染后2小时,几乎所有荧光细胞中都能检测到。这种重新定位可被环己酰亚胺阻止,但不能被盐酸胍抑制脊髓灰质炎病毒复制所阻止。与NLS融合的由三个GFP拷贝组成的蛋白质在脊髓灰质炎病毒感染后也会发生核输出。感染柯萨奇病毒B3后,EGFP-NLS也会发生类似的重新分布,感染水疱性口炎病毒后程度较轻。EGFP-NLS的流出不是由于NLS的丢失。因此,一些正链和负链RNA病毒会引发核蛋白的快速非特异性重新定位。