The Intriguing Conundrum of a Nonconserved Multifunctional Protein of Citrus Tristeza Virus That Interacts with a Viral Long Non-Coding RNA.
The Intriguing Conundrum of a Nonconserved Multifunctional Protein of Citrus Tristeza Virus That Interacts with a Viral Long Non-Coding RNA.
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DOI：
10.3390/v13112129
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发表时间：
2021-10-22
期刊：
影响因子：
--
通讯作者：
Folimonova SY
中科院分区：
文献类型：
--
作者：
Kang SH;Aknadibossian V;Kharel L;Mudiyanselage SDD;Wang Y;Folimonova SY
柑橘衰退病毒(CTV)是一种最大的非节段植物RNA病毒，它具有一种5‘端长的非编码RNA(LncRNA)，称为低分子量衰退病毒1(LMT1)。在这项研究中，我们发现p33是一种独特的病毒蛋白，在病毒感染周期中发挥多种功能，在体内和体外都能与LMT1特异性结合。这些结果是通过野生型病毒侵染背景下p33的表达或与不产生LMT1的CTV突变体一起表达，以及通过p33与LMT1在烟草叶片中的异位共表达，然后进行RNA免疫沉淀和快速扩增cDNA端分析获得的。对重组p33蛋白和体外转录的全长LMT1 RNA或其截短片段进行的进一步实验表明，p33至少与LMT1中的两个不同区域结合。据我们所知，这是第一份植物病毒蛋白与同一病毒产生的lncRNA结合的报告。对这两种病毒因子相互作用的生物学意义进行了讨论。
Citrus tristeza virus (CTV), the largest non-segmented plant RNA virus, has several peculiar features, among which is the production of a 5′-terminal long non-coding RNA (lncRNA) termed low-molecular-weight tristeza 1 (LMT1). In this study, we found that p33, a unique viral protein that performs multiple functions in the virus infection cycle, specifically binds LMT1, both in vivo and in vitro. These results were obtained through the expression of p33 under the context of the wild type virus infection or along with a mutant CTV variant that does not produce LMT1 as well as via ectopic co-expression of p33 with LMT1 in Nicotiana benthamiana leaves followed by RNA immunoprecipitation and rapid amplification of cDNA ends assays. Further experiments in which a recombinant p33 protein and an in vitro transcribed full-length LMT1 RNA or its truncated fragments were subjected to an electrophoretic mobility shift assay demonstrated that p33 binds to at least two distinct regions within LMT1. To the best of our knowledge, this is the first report of a plant virus protein binding to a lncRNA produced by the same virus. The biological significance of the interaction between these two viral factors is discussed.
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