You Jae-Hwan, Howell Gareth, Pattnaik Asit K, Osorio Fernando A, Hiscox Julian A
Institute of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds, UK.
Virology. 2008 Aug 15;378(1):34-47. doi: 10.1016/j.virol.2008.04.037. Epub 2008 Jun 12.
Porcine reproductive and respiratory syndrome virus (PRRSV), an arterivirus, in common with many other positive strand RNA viruses, encodes a nucleocapsid (N) protein which can localise not only to the cytoplasm but also to the nucleolus in virus-infected cells and cells over-expressing N protein. The dynamic trafficking of positive strand RNA virus nucleocapsid proteins and PRRSV N protein in particular between the cytoplasm and nucleolus is unknown. In this study live imaging of permissive and non-permissive cell lines, in conjunction with photo-bleaching (FRAP and FLIP), was used to investigate the trafficking of fluorescent labeled (EGFP) PRRSV-N protein. The data indicated that EGFP-PRRSV-N protein was not permanently sequestered to the nucleolus and had equivalent mobility to cellular nucleolar proteins. Further the nuclear import of N protein appeared to occur faster than nuclear export, which may account for the observed relative distribution of N protein between the cytoplasm and the nucleolus.
猪繁殖与呼吸综合征病毒(PRRSV)是一种动脉炎病毒,与许多其他正链RNA病毒一样,编码一种核衣壳(N)蛋白,该蛋白不仅可以定位于病毒感染细胞和过表达N蛋白的细胞的细胞质中,还可以定位于核仁中。正链RNA病毒核衣壳蛋白,特别是PRRSV N蛋白在细胞质和核仁之间的动态运输尚不清楚。在本研究中,使用允许和非允许细胞系的实时成像结合光漂白(FRAP和FLIP)来研究荧光标记(EGFP)的PRRSV-N蛋白的运输。数据表明,EGFP-PRRSV-N蛋白不会永久隔离在核仁中,并且与细胞核仁蛋白具有相同的流动性。此外,N蛋白的核输入似乎比核输出发生得更快,这可能解释了观察到的N蛋白在细胞质和核仁之间的相对分布情况。