Ribosomal RNA 2'-O-methylations regulate translation by impacting ribosome dynamics.
Ribosomal RNA 2'-O-methylations regulate translation by impacting ribosome dynamics.
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DOI：
10.1073/pnas.2117334119
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发表时间：
2022-03-22
影响因子：
11.1
通讯作者：
Ghalei H
中科院分区：
文献类型：
--
作者：
Khoshnevis S;Dreggors-Walker RE;Marchand V;Motorin Y;Ghalei H
RNA 化学修饰的存在早已为人所知，但其精确的分子后果仍然未知。 2'-O-甲基化是一种丰富的修饰，存在于生命各个领域的 RNA 中。核糖体 RNA (rRNA) 是一种功能重要的 RNA，经过 2'-O-甲基化的大量修饰。尽管 2'-O-甲基化在 rRNA 的重要功能区域中含量丰富，但其对核糖体活性的贡献尚不清楚。通过建立干扰rRNA 2'-O-甲基化模式的方法，我们发现rRNA 2'-O-甲基化影响核糖体的功能和保真度，并改变不同核糖体构象状态之间的平衡。我们的工作将 2'-O-甲基化与核糖体动力学联系起来，并定义了核糖体生物发生所需的一组关键 rRNA 2'-O-甲基化和其他可有可无的。核糖体的蛋白质合成对于所有细胞中的基因表达至关重要。核糖体 RNA (rRNA) 带有许多化学修饰。一组丰富的 rRNA 修饰存在于生命的各个领域，是由盒 C/D 小核仁 RNA 引导的 2'-O-甲基化，这些小核仁 RNA 是小核糖核蛋白复合物 (snoRNP) 的一部分。尽管2'-O-甲基化是核糖体正常产生所必需的，但这些修饰促进翻译的机制仍然难以捉摸。在这里，我们表明，活跃生长的酵母细胞中 C/D 盒 snoRNP 生物发生的变化导致产生具有独特翻译特性的低 2'-O-甲基化核糖体。使用 RiboMethSeq 对 2'-O-甲基化进行定量分析，我们识别了 rRNA 2'-O-甲基化模式的位点特异性扰动，并发现了正常条件下核糖体生产不需要的位点。低-2'-O-甲基化核糖体的表征揭示了显着的翻译保真度缺陷，包括移码和近同源起始密码子选择。通过rRNA结构探测，我们发现低2'-O-甲基化会影响核糖体亚基的固有动力学，并影响真核翻译起始因子1的结合，从而导致翻译缺陷。我们的数据揭示了酵母 rRNA 中不可预见的 2'-O-甲基化异质性谱，并表明 rRNA 2'-O-甲基化通过控制核糖体动力学和配体结合在调节细胞翻译中发挥重要作用。
The presence of RNA chemical modifications has long been known, but their precise molecular consequences remain unknown. 2′-O-methylation is an abundant modification that exists in RNA in all domains of life. Ribosomal RNA (rRNA) represents a functionally important RNA that is heavily modified by 2′-O-methylations. Although abundant at functionally important regions of the rRNA, the contribution of 2′-O-methylations to ribosome activities is unknown. By establishing a method to disturb rRNA 2′-O-methylation patterns, we show that rRNA 2′-O-methylations affect the function and fidelity of the ribosome and change the balance between different ribosome conformational states. Our work links 2′-O-methylation to ribosome dynamics and defines a set of critical rRNA 2′-O-methylations required for ribosome biogenesis and others that are dispensable. Protein synthesis by ribosomes is critically important for gene expression in all cells. Ribosomal RNAs (rRNAs) are marked by numerous chemical modifications. An abundant group of rRNA modifications, present in all domains of life, is 2′-O-methylation guided by box C/D small nucleolar RNAs, which are part of small ribonucleoprotein complexes (snoRNPs). Although 2′-O-methylations are required for the proper production of ribosomes, the mechanisms by which these modifications contribute to translation have remained elusive. Here, we show that a change in box C/D snoRNP biogenesis in actively growing yeast cells results in the production of hypo-2′-O-methylated ribosomes with distinct translational properties. Using RiboMethSeq for the quantitative analysis of 2′-O-methylations, we identify site-specific perturbations of the rRNA 2′-O-methylation pattern and uncover sites that are not required for ribosome production under normal conditions. Characterization of the hypo-2′-O-methylated ribosomes reveals significant translational fidelity defects, including frameshifting and near-cognate start codon selection. Using rRNA structural probing, we show that hypo-2′-O-methylation affects the inherent dynamics of the ribosomal subunits and impacts the binding of eukaryotic translation initiation factor 1, thereby causing translational defects. Our data reveal an unforeseen spectrum of 2′-O-methylation heterogeneity in yeast rRNA and suggest a significant role for rRNA 2′-O-methylation in regulating cellular translation by controlling ribosome dynamics and ligand binding.
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影响因子：
16.6
作者：
Birkedal, Ulf;Christensen-Dalsgaard, Mikkel;Nielsen, Henrik
通讯作者：
Nielsen, Henrik
影响因子：
14.9
作者：
Abou Assi H;Rangadurai AK;Shi H;Liu B;Clay MC;Erharter K;Kreutz C;Holley CL;Al-Hashimi HM
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影响因子：
13.6
作者：
Choi, Junhong;O'Loughlin, Sinead;Puglisi, Joseph D.
通讯作者：
Puglisi, Joseph D.
影响因子：
14.9
作者：
Aquino GRR;Krogh N;Hackert P;Martin R;Gallesio JD;van Nues RW;Schneider C;Watkins NJ;Nielsen H;Bohnsack KE;Bohnsack MT
通讯作者：
Bohnsack MT
影响因子：
14.9
作者：
Baudin-Baillieu A;Fabret C;Liang XH;Piekna-Przybylska D;Fournier MJ;Rousset JP
通讯作者：
Rousset JP