RNA modifications are involved in numerous biological processes. These modifications are constitutive or modulated in response to adaptive processes and can impact RNA base-pairing formation, protein recognition, RNA structure and stability. tRNAs are the most abundantly modified RNA molecules. Analysis of the roles of their modifications in response to stress, environmental changes, and infections caused by pathogens, has fueled new research areas. Nevertheless, the detection of modified nucleotides in RNAs is still a challenging task. We present here a reliable method to identify and localize tRNA modifications, which was applied to the human pathogenic bacteria, Staphyloccocus aureus. The method is based on a separation of tRNA species on a two-dimensional polyacrylamide gel electrophoresis followed by nano liquid chromatography-mass spectrometry. We provided a list of modifications mapped on 25 out of the 40 tRNA species (one isoacceptor for each amino acid). This method can be easily used to monitor the dynamics of tRNA modifications in S. aureus in response to stress adaptation and during infection of the host, a relatively unexplored field.

中文翻译：

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通过nanoLC / MSMS对金黄色葡萄球菌tRNA中的转录后修饰进行定位。

RNA修饰涉及许多生物学过程。这些修饰是组成性的或可调节的，以适应适应性过程，并可影响RNA碱基配对的形成，蛋白质识别，RNA结构和稳定性。tRNA是修饰最丰富的RNA分子。分析其修饰物对应激，环境变化和病原体引起的感染的作用，推动了新的研究领域。然而，RNA中修饰核苷酸的检测仍然是一项艰巨的任务。我们在这里提出了一种可靠的方法来鉴定和定位tRNA修饰，该修饰已应用于人类病原菌金黄色葡萄球菌。该方法基于二维聚丙烯酰胺凝胶电泳中tRNA种类的分离，然后进行纳米液相色谱-质谱分析。我们提供了在40个tRNA物种中的25个（每个氨基酸一个同一个受体）上映射的修饰列表。这种方法可以很容易地用于监测金黄色葡萄球菌中tRNA修饰的动态变化，以响应压力适应以及在宿主感染期间（一个相对未开发的领域）。