Profiling of Ribose Methylations in RNA by High-Throughput Sequencing
Profiling of Ribose Methylations in RNA by High-Throughput Sequencing
复制标题
DOI：
10.1002/anie.201408362
复制
发表时间：
2015-01-07
影响因子：
16.6
通讯作者：
Nielsen, Henrik
中科院分区：
文献类型：
--
作者：
Birkedal, Ulf;Christensen-Dalsgaard, Mikkel;Nielsen, Henrik
核糖甲基化是核糖体RNA最丰富的化学修饰，对核糖体组装和翻译的准确性至关重要。由于缺乏有效的定位方法，核糖甲基化的许多方面一直难以研究。在此，我们提出一种基于测序的方法（RiboMeth - seq）及其在酵母核糖体中的应用，酵母核糖体是目前研究最为深入的真核模式系统。我们展示了对已知以及新的修饰的检测，揭示了部分修饰以及修饰事件之间意想不到的关联，并确定了核糖体生物发生过程中几个位点的修饰顺序。令人惊讶的是，该方法还提供了关于其他一部分修饰的信息。因此，RiboMeth - seq能够详细评估RNA修饰在细胞中最复杂的分子机器中的重要性。RiboMeth - seq可适用于其他RNA类型，例如mRNA，以揭示涉及RNA修饰的新的生物学现象。
Ribose methylations are the most abundant chemical modifications of ribosomal RNA and are critical for ribosome assembly and fidelity of translation. Many aspects of ribose methylations have been difficult to study due to lack of efficient mapping methods. Here, we present a sequencing-based method (RiboMeth-seq) and its application to yeast ribosomes, presently the best-studied eukaryotic model system. We demonstrate detection of the known as well as new modifications, reveal partial modifications and unexpected communication between modification events, and determine the order of modification at several sites during ribosome biogenesis. Surprisingly, the method also provides information on a subset of other modifications. Hence, RiboMeth-seq enables a detailed evaluation of the importance of RNA modifications in the cells most sophisticated molecular machine. RiboMeth-seq can be adapted to other RNA classes, for example, mRNA, to reveal new biology involving RNA modifications.