Fujisaki Koki, Ishikawa Masayuki
Plant-Microbe Interactions Research Unit, National Institute of Agrobiological Sciences, 2-1-2 Kan-non-dai, Tsukuba 305-8602, Japan.
Virology. 2008 Oct 25;380(2):402-11. doi: 10.1016/j.virol.2008.07.033. Epub 2008 Aug 31.
The genomic RNAs of positive-strand RNA viruses carry RNA elements that play positive, or in some cases, negative roles in virus multiplication by interacting with viral and cellular proteins. In this study, we purified Arabidopsis thaliana proteins that specifically bind to 5' or 3' terminal regions of tomato mosaic virus (ToMV) genomic RNA, which contain important regulatory elements for translation and RNA replication, and identified these proteins by mass spectrometry analyses. One of these host proteins, named BTR1, harbored three heterogeneous nuclear ribonucleoprotein K-homology RNA-binding domains and preferentially bound to RNA fragments that contained a sequence around the initiation codon of the 130K and 180K replication protein genes. The knockout and overexpression of BTR1 specifically enhanced and inhibited, respectively, ToMV multiplication in inoculated A. thaliana leaves, while such effect was hardly detectable in protoplasts. These results suggest that BTR1 negatively regulates the local spread of ToMV.
正链RNA病毒的基因组RNA携带RNA元件,这些元件通过与病毒和细胞蛋白相互作用,在病毒增殖中发挥正向作用,或在某些情况下发挥负向作用。在本研究中,我们纯化了与番茄花叶病毒(ToMV)基因组RNA的5'或3'末端区域特异性结合的拟南芥蛋白,这些区域包含翻译和RNA复制的重要调控元件,并通过质谱分析鉴定了这些蛋白。其中一种宿主蛋白名为BTR1,它含有三个不均一核核糖核蛋白K同源RNA结合结构域,并优先结合包含130K和180K复制蛋白基因起始密码子周围序列的RNA片段。BTR1的敲除和过表达分别特异性增强和抑制了接种的拟南芥叶片中ToMV的增殖,而在原生质体中几乎检测不到这种效应。这些结果表明,BTR1对ToMV的局部传播起负调控作用。