The Generation of Turnip Crinkle Virus-Like Particles in Plants by the Transient Expression of Wild-Type and Modified Forms of Its Coat Protein.
The Generation of Turnip Crinkle Virus-Like Particles in Plants by the Transient Expression of Wild-Type and Modified Forms of Its Coat Protein.
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DOI：
10.3389/fpls.2015.01138
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发表时间：
2015
影响因子：
5.6
通讯作者：
Lomonossoff GP
中科院分区：
文献类型：
--
作者：
Saunders K;Lomonossoff GP
芜菁皱缩病毒（Turnip crinkle virus，TCV）是番茄丛矮病毒科（Tombusviridae）的一种花叶病毒属（Carmovirus）病毒，其基因组由一个单义RNA分子组成，该RNA分子包埋在一个二十面体颗粒中，该颗粒由180个拷贝的单一类型的外壳蛋白组成。我们已经采用了CPMV-HT瞬时表达系统来研究TCV样颗粒的形成后，表达的野生型外壳蛋白或修改后的形式，它包含删除和/或添加。外壳蛋白在植物中的瞬时表达导致衣壳结构的形成，所述衣壳结构在形态上类似于TCV病毒体（T = 3结构），但衣壳化异质细胞RNA，而不是特异性TCV外壳蛋白信使RNA。氨基末端缺失形式的外壳蛋白的表达导致形成不含RNA的较小T = 1结构。利用TCV作为载体的外源蛋白的颗粒表面上的介绍的可能性也探讨了通过融合的序列的GFP的外壳蛋白的C-末端。外壳蛋白-GFP杂交体的表达允许VLP的形成，但与用未修饰的外壳蛋白获得的产量相比，颗粒的产量减少。我们的研究结果证实了N-末端的外壳蛋白的RNA的bissidation的重要性，并表明，外壳蛋白的外部P结构域在颗粒形成中起着关键作用。
Turnip crinkle virus (TCV), a member of the genus carmovirus of the Tombusviridae family, has a genome consisting of a single positive-sense RNA molecule that is encapsidated in an icosahedral particle composed of 180 copies of a single type of coat protein. We have employed the CPMV-HT transient expression system to investigate the formation of TCV-like particles following the expression of the wild-type coat protein or modified forms of it that contain either deletions and/or additions. Transient expression of the coat protein in plants results in the formation of capsid structures that morphologically resemble TCV virions (T = 3 structure) but encapsidate heterogeneous cellular RNAs, rather than the specific TCV coat protein messenger RNA. Expression of an amino-terminal deleted form of the coat protein resulted in the formation of smaller T = 1 structures that are free of RNA. The possibility of utilizing TCV as a carrier for the presentation of foreign proteins on the particle surface was also explored by fusing the sequence of GFP to the C-terminus of the coat protein. The expression of coat protein-GFP hybrids permitted the formation of VLPs but the yield of particles is diminished compared to the yield obtained with unmodified coat protein. Our results confirm the importance of the N-terminus of the coat protein for the encapsidation of RNA and show that the coat protein's exterior P domain plays a key role in particle formation.