Two orthogonal cleavages separate subunit RNAs in mouse ribosome biogenesis.
Two orthogonal cleavages separate subunit RNAs in mouse ribosome biogenesis.
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DOI：
10.1093/nar/gku787
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发表时间：
2014
影响因子：
14.9
通讯作者：
Pestov DG
中科院分区：
文献类型：
--
作者：
Wang M;Anikin L;Pestov DG
核糖体的生物发生是一个动态的多步骤过程，其中的许多特征仍未得到完整的记载。在这里，我们展示了这一途径的变化可以通过一组前rrna比率的图形来捕获和注释，我们称之为多前体比率分析（RAMP）的技术。我们发现，敲除核糖体合成因子会产生一种特征的RAMP谱，该谱在耗尽水平范围内表现出一致性。这有助于推断受影响的步骤，简化比较分析。我们应用RAMP研究了小鼠内部转录间隔1 （ITS1）中pre-rRNA转录物的核内溶裂解如何受到小核糖体亚基（Rcl1, Fcf1/Utp24, Utp23）和大亚基（Pes1, Nog1）成熟所需因子的消耗的影响。这些数据表明，亚基的早期成熟通过刺激ITS1近端切割，触发其从常见的pre-rRNA转录物中释放出来。我们还发现ITS1 3 '区pre-rRNA的分裂在成年小鼠组织和静止细胞中普遍存在，就像在人类细胞中一样。我们提出了哺乳动物核糖体合成过程中亚基分离的模型，并讨论了其对理解前rrna加工途径的意义。
Ribosome biogenesis is a dynamic multistep process, many features of which are still incompletely documented. Here, we show that changes in this pathway can be captured and annotated by means of a graphic set of pre-rRNA ratios, a technique we call Ratio Analysis of Multiple Precursors (RAMP). We find that knocking down a ribosome synthesis factor produces a characteristic RAMP profile that exhibits consistency across a range of depletion levels. This facilitates the inference of affected steps and simplifies comparative analysis. We applied RAMP to examine how endonucleolytic cleavages of the mouse pre-rRNA transcript in the internal transcribed spacer 1 (ITS1) are affected by depletion of factors required for maturation of the small ribosomal subunit (Rcl1, Fcf1/Utp24, Utp23) and the large subunit (Pes1, Nog1). The data suggest that completion of early maturation in a subunit triggers its release from the common pre-rRNA transcript by stimulating cleavage at the proximal site in ITS1. We also find that splitting of pre-rRNA in the 3′ region of ITS1 is prevalent in adult mouse tissues and quiescent cells, as it is in human cells. We propose a model for subunit separation during mammalian ribosome synthesis and discuss its implications for understanding pre-rRNA processing pathways.
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