Department of Cellular and Molecular Medicine, University of Copenhagen, Denmark.
Department of Cellular and Molecular Medicine, University of Copenhagen, Denmark.
Methods. 2019 Mar 1;156:5-15. doi: 10.1016/j.ymeth.2018.11.017. Epub 2018 Nov 29.
Ribose methylation is one of the most abundant RNA modifications and is found in all domains of life and all major classes of RNA (rRNA, tRNA, and mRNA). Ribose methylations are introduced by stand-alone enzymes or by generic enzymes guided to the target by small RNA guides. Recent years have seen the development of several sequencing-based methods for RNA modifications relying on different principles. In this review, we compare mapping and quantitation studies of ribose methylations from yeast and human culture cells. The emphasis is on ribosomal RNA for which the results can be compared to results from RNA fingerprinting and mass spectrometry. One sequencing approach is consistent with these methods and paints a conservative picture of rRNA modifications. Other approaches detect many more sites. Similar discrepancies are found in measurements of modification stoichiometry. The results are discussed in relation to the more challenging task of mapping ribose methylations in mRNA.
核糖甲基化是最丰富的 RNA 修饰之一,存在于所有生命领域和所有主要类型的 RNA(rRNA、tRNA 和 mRNA)中。核糖甲基化是由独立的酶或由小 RNA 引导的通用酶引入到靶标上的。近年来,已经开发出了几种基于测序的 RNA 修饰方法,这些方法依赖于不同的原理。在这篇综述中,我们比较了来自酵母和人类培养细胞的核糖甲基化的图谱和定量研究。重点是核糖体 RNA,其结果可以与 RNA 指纹图谱和质谱法的结果进行比较。一种测序方法与这些方法一致,并描绘了 rRNA 修饰的保守图景。其他方法检测到的位点要多得多。在修饰化学计量的测量中也发现了类似的差异。结果与在 mRNA 中绘制核糖甲基化图谱这一更具挑战性的任务有关。