Systematic mapping of rRNA 2'-O methylation during frog development and involvement of the methyltransferase Fibrillarin in eye and craniofacial development in Xenopus laevis.
Systematic mapping of rRNA 2'-O methylation during frog development and involvement of the methyltransferase Fibrillarin in eye and craniofacial development in Xenopus laevis.
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DOI：
10.1371/journal.pgen.1010012
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发表时间：
2022-01
期刊：
影响因子：
4.5
通讯作者：
Lafontaine DLJ
中科院分区：
文献类型：
--
作者：
Delhermite J;Tafforeau L;Sharma S;Marchand V;Wacheul L;Lattuca R;Desiderio S;Motorin Y;Bellefroid E;Lafontaine DLJ
核糖体是负责蛋白质生产的重要纳米机器。虽然核糖体存在于每个活细胞中，但核糖体生物合成功能障碍疾病（称为核糖体病）会特异性地影响特定组织。在这里，我们评估盒C/D snoRNA相关的核糖体RNA甲基转移酶原纤蛋白（FBL）在非洲爪蟾早期胚胎发育的重要性。我们报告说，在发育中的胚胎，神经板，神经嵴细胞（NCC），和NCC衍生物丰富的fbl成绩单。Fbl敲除导致影响眼睛和颅面骨骼的显著形态学缺陷，这是由于大量p53依赖性细胞凋亡引起的NCC存活的缺乏。Fbl是有效的pre-rRNA加工和18 S rRNA产生所必需的，这解释了早期发育缺陷。使用RiboMethSeq，我们系统地重新研究了X中的核糖体RNA 2 '-O甲基化。laevis，确认了所有89个先前定位的位点，并确定了18 S和28 S rRNA中的15个新的推定位置。23个位置，包括10个新的，通过低dNTP引物延伸进行正交验证。利用生物信息学技术对黄曲藻进行了筛选。laevis转录组揭示了所有甲基化位置的候选盒C/D snoRNA。在单个胚胎的6个发育阶段的2 '-O甲基化的定位表明，在发育过程中的特定位置的甲基化减少的趋势。我们的结论是，在早期非洲爪蟾胚胎原纤蛋白敲低导致功能性核糖体亚基的生产减少，从而损害NCC的形成和迁移。核糖体是负责所有细胞中蛋白质生产的重要纳米机器。核糖体病是由于核糖体蛋白或核糖体组装因子的突变导致的不适当的核糖体形成引起的疾病。这些疾病主要影响大脑和血液，目前还不清楚像核糖体形成这样的一般过程的故障如何导致组织特异性疾病。在这里，我们已经研究了如何原纤蛋白，一种酶，修改核糖体RNA通过添加甲基基团在特定的网站，影响早期胚胎发育的青蛙非洲爪蟾。我们已经揭示了它在细胞成熟过程中的重要性，这些细胞形成了一种称为神经嵴的胚胎结构。原纤蛋白耗竭导致眼睛尺寸减小和头部形状异常，使人联想到其他病症，如Treacher柯林斯综合征。在分子上，所观察到的表型可解释为通过抑制某些前rRNA加工步骤触发的p53依赖性程序性细胞死亡增加。我们对发育过程中核糖体RNA 2 '-O甲基化库的系统研究进一步揭示了发育后期的低修饰，这可能在涉及组成不同的核糖体差异翻译的晚期发育过渡中发挥作用。
Ribosomes are essential nanomachines responsible for protein production. Although ribosomes are present in every living cell, ribosome biogenesis dysfunction diseases, called ribosomopathies, impact particular tissues specifically. Here, we evaluate the importance of the box C/D snoRNA-associated ribosomal RNA methyltransferase fibrillarin (Fbl) in the early embryonic development of Xenopus laevis. We report that in developing embryos, the neural plate, neural crest cells (NCCs), and NCC derivatives are rich in fbl transcripts. Fbl knockdown leads to striking morphological defects affecting the eyes and craniofacial skeleton, due to lack of NCC survival caused by massive p53-dependent apoptosis. Fbl is required for efficient pre-rRNA processing and 18S rRNA production, which explains the early developmental defects. Using RiboMethSeq, we systematically reinvestigated ribosomal RNA 2’-O methylation in X. laevis, confirming all 89 previously mapped sites and identifying 15 novel putative positions in 18S and 28S rRNA. Twenty-three positions, including 10 of the new ones, were validated orthogonally by low dNTP primer extension. Bioinformatic screening of the X. laevis transcriptome revealed candidate box C/D snoRNAs for all methylated positions. Mapping of 2’-O methylation at six developmental stages in individual embryos indicated a trend towards reduced methylation at specific positions during development. We conclude that fibrillarin knockdown in early Xenopus embryos causes reduced production of functional ribosomal subunits, thus impairing NCC formation and migration. Ribosomes are essential nanomachines responsible for protein production in all cells. Ribosomopathies are diseases caused by improper ribosome formation due to mutations in ribosomal proteins or ribosome assembly factors. Such diseases primarily affect the brain and blood, and it is unclear how malfunctioning of a process as general as ribosome formation can lead to tissue-specific diseases. Here we have examined how fibrillarin, an enzyme which modifies ribosomal RNA by adding methyl groups at specific sites, affects early embryonic development in the frog Xenopus laevis. We have revealed its importance in the maturation of cells forming an embryonic structure called the neural crest. Fibrillarin depletion leads to reduced eye size and abnormal head shape, reminiscent of other conditions such as Treacher Collins syndrome. Molecularly, the observed phenotypes are explainable by increased p53-dependent programmed cell death triggered by inhibition of certain pre-rRNA processing steps. Our systematic investigation of the ribosomal RNA 2’-O methylation repertoire across development has further revealed hypomodification at a late stage of development, which might play a role in late developmental transitions involving differential translation by compositionally different ribosomes.
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在人rRNA中2'-O-ME的分析揭示了一部分部分修饰的位置，并提供了核糖体异质性的证据。
影响因子：
14.9
作者：
Krogh N;Jansson MD;Häfner SJ;Tehler D;Birkedal U;Christensen-Dalsgaard M;Lund AH;Nielsen H
通讯作者：
Nielsen H
影响因子：
7.3
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影响因子：
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作者：
Griffin JN;Sondalle SB;Del Viso F;Baserga SJ;Khokha MK
通讯作者：
Khokha MK
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：
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通讯作者：
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