Pooggin Mikhail M, Fütterer Johannes, Hohn Thomas
Institute of Botany, University of Basel, Basel, Switzerland.
PLoS One. 2008 Feb 20;3(2):e1650. doi: 10.1371/journal.pone.0001650.
Cauliflower mosaic virus (CaMV) and Rice tungro bacilliform virus (RTBV) belong to distinct genera of pararetroviruses infecting dicot and monocot plants, respectively. In both viruses, polycistronic translation of pregenomic (pg) RNA is initiated by shunting ribosomes that bypass a large region of the pgRNA leader with several short (s)ORFs and a stable stem-loop structure. The shunt requires translation of a 5'-proximal sORF terminating near the stem. In CaMV, mutations knocking out this sORF nearly abolish shunting and virus viability.
METHODOLOGY/PRINCIPAL FINDINGS: Here we show that two distant regions of the CaMV leader that form a minimal shunt configuration comprising the sORF, a bottom part of the stem, and a shunt landing sequence can be replaced by heterologous sequences that form a structurally similar configuration in RTBV without any dramatic effect on shunt-mediated translation and CaMV infectivity. The CaMV-RTBV chimeric leader sequence was largely stable over five viral passages in turnip plants: a few alterations that did eventually occur in the virus progenies are indicative of fine tuning of the chimeric sequence during adaptation to a new host.
CONCLUSIONS/SIGNIFICANCE: Our findings demonstrate cross-species functionality of pararetroviral cis-elements driving ribosome shunting and evolutionary conservation of the shunt mechanism. We are grateful to Matthias Müller and Sandra Pauli for technical assistance. This work was initiated at Friedrich Miescher Institute (Basel, Switzerland). We thank Prof. Thomas Boller for hosting the group at the Institute of Botany.
花椰菜花叶病毒(CaMV)和水稻东格鲁杆状病毒(RTBV)分别属于感染双子叶植物和单子叶植物的不同副逆转录病毒属。在这两种病毒中,前基因组(pg)RNA的多顺反子翻译是由跳跃核糖体启动的,这些核糖体绕过pgRNA前导序列的一大段区域,该区域包含几个短(s)开放阅读框(ORF)和一个稳定的茎环结构。这种跳跃需要翻译一个在茎附近终止的5'-近端sORF。在CaMV中,敲除这个sORF的突变几乎消除了跳跃和病毒活力。
方法/主要发现:在这里,我们表明,CaMV前导序列的两个远距离区域形成了一个最小的跳跃结构,包括sORF、茎的底部和一个跳跃着陆序列,可以被RTBV中形成结构相似结构的异源序列所取代,而对跳跃介导的翻译和CaMV感染性没有任何显著影响。CaMV-RTBV嵌合前导序列在芜菁植物的五次病毒传代中基本稳定:病毒后代中最终出现的一些变化表明在适应新宿主的过程中对嵌合序列进行了微调。
结论/意义:我们的发现证明了驱动核糖体跳跃的副逆转录病毒顺式元件的跨物种功能以及跳跃机制的进化保守性。我们感谢马蒂亚斯·米勒和桑德拉·保利提供的技术帮助。这项工作始于弗里德里希·米歇尔研究所(瑞士巴塞尔)。我们感谢托马斯·博勒教授在植物研究所接待该研究小组。