Rainsford Edward W, McCrae Malcolm A
Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, United Kingdom.
Virus Res. 2007 Dec;130(1-2):193-201. doi: 10.1016/j.virusres.2007.06.011. Epub 2007 Jul 20.
The 12kDa non-structural protein 6 (NSP6) is the least studied of the rotavirus proteins. In an attempt to further characterize this protein mono-specific antisera was generated using purified protein expressed in E. coli. Pulse/chase radio-labeling of virus infected cells was used to show that it is expressed at a steady but low rate throughout the virus replication cycle. In contrast to the other rotavirus non-structural proteins, NSP6 was found to have a high rate of turnover, being completely degraded within 2h of synthesis. NSP6 tagged with GFP was used to probe the intracellular distribution of the protein, perinuclear aggregates were observed in the cytoplasm of transfected cells. Following virus infection of these transfected cells the aggregates were seen to redistribute to the viroplasms. Consistent with its localization to the site of viral genome replication and packaging, NSP6 was found to be a sequence independent nucleic acid binding protein, with similar affinities for ssRNA and dsRNA.
12千道尔顿非结构蛋白6（NSP6）是轮状病毒蛋白中研究最少的一种。为了进一步表征该蛋白，利用在大肠杆菌中表达的纯化蛋白制备了单特异性抗血清。通过对病毒感染细胞进行脉冲/追踪放射性标记，结果表明在整个病毒复制周期中，NSP6均以稳定但较低的速率表达。与其他轮状病毒非结构蛋白不同，NSP6的周转速度很快，在合成后2小时内就会完全降解。用绿色荧光蛋白（GFP）标记的NSP6用于探测该蛋白在细胞内的分布，在转染细胞的细胞质中观察到核周聚集物。这些转染细胞受到病毒感染后，聚集物会重新分布到病毒工厂。与它定位于病毒基因组复制和包装位点一致，NSP6被发现是一种不依赖序列的核酸结合蛋白，对单链RNA和双链RNA具有相似的亲和力。
