中文摘要：N-6甲基腺苷（m(6)A）是许多物种中比较普遍的RNA修饰。m(6)A甲基化水平异常可导致RNA功能障碍并可引起疾病。烟草花叶病毒（TMV）是农业植物中最具破坏性的病毒之一。TMV寄主很多，包括烟草和茄科的其他一些成员。然而，尚不清楚TMV导致的异常生长是否与m(6)A水平相关。本研究采用超高效液相色谱-高分辨率串联质谱（UHPLC-HR - MS / MS）方法，快速准确地分析烟叶中腺苷（A），胞苷（C），鸟苷（ G），尿苷（U）和m（6）A的含量，并利用m（6）A/G比评价m（6）A水平。研究结果显示，TMV降低了m（6）A水平。此外，蛋白质序列比对揭示了人ALKBH5，拟南芥（NP_001031793）和烟草（Nicotiana sylvestris）（XP_009800010）之间的部分同源性。在感染TMV的普通烟草中，潜在的去甲基化酶XM_009801708的基因表达水平在第14天和第21天增加，而所有潜在的甲基化酶减少。烟草mRNA中 m(6)A的可逆修饰可能代表了一种关于TMV的新的表观遗传机制。
外文摘要：Background: N-6-methyl-adenosine (m(6)A) is a prevalent RNA modification in many species. Abnormal m(6)A methylation levels can lead to RNA dysfunction and can cause diseases. Tobacco mosaic virus (TMV) is one of the most devastating viruses for agricultural plants. It has many hosts, particularly including tobacco and other members the family Solanaceae. However, it remains unclear whether the abnormal growth induced by TMV is associated with the m(6)A level.
Methods: A rapid and accurate analytical method using ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry (UHPLC-HR - MS/MS) was developed to analyse the adenosine (A), cytidine (C), guanosine (G), uridine (U), and m(6)A contents in the tobacco leaf, and the m(6)A/G ratio was used to evaluate the m(6)A level. Subsequent protein sequence alignments were used to find the potential methylases and demethylases in Nicotiana tabacum (N. tabacum). Finally, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was used to analyse the gene expression levels of the potential methylases and demethylases in the N. tabacum leaf.
Results: The results showed that TMV reduced the m(6)A level. Moreover, protein sequence alignments revealed partial homology among human ALKBH5, Arabidopsis (NP_001031793), and Nicotiana sylvestris (XP_009800010). The gene expression level of the potential demethylase XM_009801708 increased at 14 and 21 days in N. tabacum infected with TMV, whereas all of the potential methylases decreased.
Conclusions: The reversible m(6)A modification in N. tabacum mRNA might represent a novel epigenetic mechanism involved in TMV.
外文关键词：N. tabacum; TMV; m(6)A; UHPLC-HR - MS/MS; RT-qPCR
作者：Li, Zhurui; Shi, Jing; Yu, Lu; 等
作者单位：贵州大学
期刊名称：VIROLOGY JOURNAL
期刊影响因子：2.465
出版年份：2018
出版刊次：5
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