The initial U3 snoRNA:pre-rRNA base pairing interaction required for pre-18S rRNA folding revealed by in vivo chemical probing.
The initial U3 snoRNA:pre-rRNA base pairing interaction required for pre-18S rRNA folding revealed by in vivo chemical probing.
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DOI：
10.1093/nar/gkr044
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发表时间：
2011-07
影响因子：
14.9
通讯作者：
Baserga SJ
中科院分区：
文献类型：
--
作者：
Dutca LM;Gallagher JE;Baserga SJ
核仁中核糖体亚基的合成是一个保守的、必需的过程，其导致具有精确加工和折叠的rRNA与核糖体蛋白组装的细胞质核糖体。有人提出，但从未直接证明，U3小核仁RNA（snoRNA），核糖体生物合成所需的核仁组分，是前18 S rRNA折叠的伴侣。为了验证这一点，我们使用硫酸二甲酯进行体内化学探测，以检测酵母（Saccharomyces cerevisiae）遗传操作后前rRNA结构的变化。基于核苷酸反应性的变化，我们发现U3 snoRNA确实是前18 S rRNA折叠所必需的。此外，我们检测到一个新的基本碱基配对相互作用，这可能是最初的锚招募U3 snoRNA的前rRNA，是一个先决条件，为后续的相互作用，并需要为小亚基加工体的形成。参与这种初始碱基配对相互作用的前体rRNA的5′-ETS核苷酸的取代是致命的，但当互补的U3 snoRNA表达时，生长恢复。通过碱基配对，U3 snoRNP及其相关蛋白是协调导致小核糖体亚基组装的前体rRNA折叠所需机制的一部分。
The synthesis of ribosomal subunits in the nucleolus is a conserved, essential process that results in cytoplasmic ribosomes with precisely processed and folded rRNAs assembled with ribosomal proteins. It has been proposed, but never directly demonstrated, that the U3 small nucleolar RNA (snoRNA), a nucleolar component required for ribosome biogenesis, is a chaperone for pre-18S rRNA folding. To test this, we used in vivo chemical probing with dimethyl sulfate to detect changes in pre-rRNA structure upon genetic manipulation of the yeast, Saccharomyces cerevisiae. Based on changes in nucleotide reactivity, we found that the U3 snoRNA is indeed required for folding of the pre-18S rRNA. Furthermore, we detected a new essential base pairing interaction that is likely the initial anchor that recruits the U3 snoRNA to the pre-rRNA, is a prerequisite for the subsequent interactions, and is required for the small subunit processome formation. Substitution of the 5′-ETS nucleotides of the pre-rRNA involved in this initial base pairing interaction is lethal, but growth is restored when a complementary U3 snoRNA is expressed. The U3 snoRNP, via base pairing, and its associated proteins, are part of the required machinery that orchestrates the folding of pre-rRNA that results in the assembly of the small ribosomal subunit.
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