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首页> 外文期刊>Nucleic acids research >Characterization of the RNA motif responsible for the specific interaction of potato spindle tuber viroid RNA (PSTVd) and the tomato protein Virp1
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Characterization of the RNA motif responsible for the specific interaction of potato spindle tuber viroid RNA (PSTVd) and the tomato protein Virp1

机译：负责马铃薯纺锤块茎类病毒RNA（PSTVd）与番茄蛋白Virp1特异性相互作用的RNA基序的表征

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摘要

Viroids are small non‐coding parasitic RNAs that are able to infect their host plants systemically. This circular naked RNA makes use of host proteins to accomplish its proliferation. Here we analyze the specific binding of the tomato protein Virp1 to the terminal right domain of potato spindle tuber viroid RNA (PSTVd). We find that two asymmetric internal loops within the PSTVd (+) RNA, each composed of the sequence elements 5′‐ACAGG and CUCUUCC‐5′, are responsible for the specific RNA–protein interaction. In view of the nucleotide composition we call this structural element an ‘RY motif'. The RY motif located close to the terminal right hairpin loop of the PSTVd secondary structure has an ～5‐fold stronger binding affinity than the more centrally located RY motif. Simultaneous sequence alterations in both RY motifs abolished the specific binding to Virp1. Mutations in any of the two RY motifs resulted in non‐infectious viroid RNA, with the exception of one case, where reversion to sequence wild type took place. In contrast, the simultaneous exchange of two nucleotides within the terminal right hairpin loop of PSTVd had only moderate influence on the binding to Virp1. This variant was infectious and sequence changes were maintained in the progeny. The relevance of the phylogenetic conservation of the RY motif, and sequence elements therein, amongst various genera of the family Pospiviroidae is discussed.
机译：病毒是小的非编码寄生RNA，能够系统感染其宿主植物。该环状裸RNA利用宿主蛋白来完成其增殖。在这里，我们分析了番茄蛋白Virp1与马铃薯纺锤块茎类病毒RNA（PSTVd）末端右域的特异性结合。我们发现PSTVd（+）RNA中有两个不对称内部环，每个环由5'-ACAGG和CUCUUCC-5'序列元素组成，负责特定的RNA-蛋白相互作用。考虑到核苷酸的组成，我们称这种结构元件为“ RY基序”。靠近PSTVd二级结构的末端右发夹环的RY基序比位于中心位置的RY基序具有约5倍的结合亲和力。两个RY基序中的同时序列更改取消了与Virp1的特异性结合。两个RY基序中的任何一个突变均导致非感染性类病毒RNA，只有一种情况例外，即发生了向野生型序列的回复。相反，PSTVd末端右发夹环内两个核苷酸的同时交换仅对与Virp1的结合产生中等影响。该变体具有感染性，并且后代中的序列改变得以维持。讨论了RY基序和其中的序列元件的系统发育保守性的相关性，该基因在马铃薯假单胞菌科的各个属中。

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