Schaad M C, Anderberg R J, Carrington J C
Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340, USA.
Virology. 2000 Aug 1;273(2):300-6. doi: 10.1006/viro.2000.0416.
The NIa protein of potyviruses provides VPg and proteolytic functions during virus replication. It has also been shown to confer host genotype-specific movement functions in plants. Specifically, NIa from tobacco etch virus (TEV)-Oxnard, but not from most other strains, confers the ability to move long distances in Nicotiana tabacum cultivar "V-20." This led to the hypothesis that all or part of NIa may interact with one or more cellular factors. To identify cellular proteins that interact with NIa in a host- or strain-specific manner, a yeast two-hybrid search of a tomato cDNA library was done. Ten proteins that interacted with NIa were recovered, with translation initiation factor eIF4E being by far the most common protein identified. Interaction of eIF4E with NIa was shown to be TEV strain-specific. eIF4E from both tomato and tobacco interacted well with NIa from the HAT strain, but not from the Oxnard strain. However, using chimeric NIa proteins, the determinant for systemic infection of V20 plants was found to be genetically distinct from the determinant controlling eIF4E interaction. In TEV-eIF4E coexpression experiments, evidence suggesting that eIF4E provides a positive effect on genome amplification was obtained.
马铃薯Y病毒属病毒的NIa蛋白在病毒复制过程中发挥VPg和蛋白水解功能。研究还表明,它在植物中赋予宿主基因型特异性的移动功能。具体而言,烟草蚀纹病毒(TEV)-奥克斯纳德株系的NIa蛋白能使病毒在烟草品种“V-20”中长距离移动,而大多数其他株系的NIa蛋白则不能。这引发了一个假设,即NIa的全部或部分可能与一种或多种细胞因子相互作用。为了鉴定以宿主或株系特异性方式与NIa相互作用的细胞蛋白,对番茄cDNA文库进行了酵母双杂交筛选。共筛选到10种与NIa相互作用的蛋白,其中翻译起始因子eIF4E是最常见的蛋白。结果表明,eIF4E与NIa的相互作用具有TEV株系特异性。番茄和烟草中的eIF4E都能与HAT株系的NIa良好互作,但不能与奥克斯纳德株系的NIa互作。然而,通过嵌合NIa蛋白研究发现,V20植物系统感染的决定因素在遗传上与控制eIF4E相互作用的决定因素不同。在TEV-eIF4E共表达实验中,获得了表明eIF4E对基因组扩增有积极作用的证据。