A hierarchical model for assembly of eukaryotic 60S ribosomal subunit domains.
A hierarchical model for assembly of eukaryotic 60S ribosomal subunit domains.
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DOI：
10.1101/gad.228825.113
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发表时间：
2014-01-15
影响因子：
10.5
通讯作者：
Woolford JL Jr
中科院分区：
文献类型：
--
作者：
Gamalinda M;Ohmayer U;Jakovljevic J;Kumcuoglu B;Woolford J;Mbom B;Lin L;Woolford JL Jr
尽管具有真核生物大核糖体亚基的结构，但这些核糖核蛋白复合物如何在活细胞中构建尚不清楚。在这里，Gamalinda等人使用蛋白质组学来测定在60 S组装中起作用的核糖体蛋白耗尽后前核糖体中核糖体蛋白和组装因子水平的变化。结果表明，真核生物60 S核糖体亚基是以分级方式组装的。这项研究进一步揭示了细菌和真核生物大核糖体亚基之间的显着差异和相似性，为RNA蛋白颗粒的进化提供了见解。尽管真核生物大核糖体亚基具有高分辨率结构，但这些核糖核蛋白复合物如何在活细胞中构建仍不清楚。然而，知道核糖体蛋白与核糖体RNA（rRNA）相互作用的位置提供了一个战略平台，以调查60 S亚基生物发生的空间和时间方面之间的联系。我们以前发现，单个酵母大亚基核糖体蛋白（RPLs）在前体rRNA（pre-rRNA）加工中的功能与它们在成熟60 S亚基结构中的位置相关。这一观察结果表明，60 S亚基的形成是有顺序的。为了测试这个模型，我们使用蛋白质组学的方法来测定核糖体蛋白和装配因子的水平在前核糖体的RPL功能时，在早期，中期和晚期步骤的前60 S装配耗尽的变化。我们的研究结果表明，真核细胞60 S核糖体亚基的结构域形成的层次方式。组装开始于凸形溶剂侧，接着是多肽出口通道、亚基间侧，最后是中央突起。该模型为真核生物60 S亚基的顺序组装提供了一个初始范例。我们的研究结果揭示了细菌和真核生物大核糖体亚基组装之间的显着差异和相似性，为这些RNA蛋白颗粒的进化提供了见解。
Despite having structures for eukaryotic large ribosomal subunits, it has been unclear how these ribonucleoprotein complexes are constructed in living cells. Here, Gamalinda et al. used proteomics to assay changes in the levels of ribosomal protein and assembly factors in preribosomes upon depletion of ribosomal proteins that function in 60S assembly. The results show that the eukaryotic 60S ribosomal subunits are assembled in a hierarchical fashion. This study further reveals striking differences and similarities between bacterial and eukaryotic large ribosomal subunits, providing insights into the evolution of RNA–protein particles. Despite having high-resolution structures for eukaryotic large ribosomal subunits, it remained unclear how these ribonucleoprotein complexes are constructed in living cells. Nevertheless, knowing where ribosomal proteins interact with ribosomal RNA (rRNA) provides a strategic platform to investigate the connection between spatial and temporal aspects of 60S subunit biogenesis. We previously found that the function of individual yeast large subunit ribosomal proteins (RPLs) in precursor rRNA (pre-rRNA) processing correlates with their location in the structure of mature 60S subunits. This observation suggested that there is an order by which 60S subunits are formed. To test this model, we used proteomic approaches to assay changes in the levels of ribosomal proteins and assembly factors in preribosomes when RPLs functioning in early, middle, and late steps of pre-60S assembly are depleted. Our results demonstrate that structural domains of eukaryotic 60S ribosomal subunits are formed in a hierarchical fashion. Assembly begins at the convex solvent side, followed by the polypeptide exit tunnel, the intersubunit side, and finally the central protuberance. This model provides an initial paradigm for the sequential assembly of eukaryotic 60S subunits. Our results reveal striking differences and similarities between assembly of bacterial and eukaryotic large ribosomal subunits, providing insights into how these RNA–protein particles evolved.
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