Visualizing the Assembly Pathway of Nucleolar Pre-60S Ribosomes.
Visualizing the Assembly Pathway of Nucleolar Pre-60S Ribosomes.
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DOI：
10.1016/j.cell.2017.11.039
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发表时间：
2017-12-14
期刊：
影响因子：
64.5
通讯作者：
Beckmann R
中科院分区：
文献类型：
--
作者：
Kater L;Thoms M;Barrio-Garcia C;Cheng J;Ismail S;Ahmed YL;Bange G;Kressler D;Berninghausen O;Sinning I;Hurt E;Beckmann R
真核60S核糖体亚基由3个rna和约50个核糖体蛋白组成。它们形成的最初步骤发生在核仁中，但是，由于缺乏结构信息，这一过程知之甚少。利用低温电镜技术，我们以3.3 Å至4.5 Å的分辨率解析了早期60S生物发生中间体的结构，从而深入了解了它们的顺序折叠和组装途径。除了揭示不同的未成熟rRNA构象外，我们还绘制了六种不同组装状态下的25个组装因子。值得注意的是，Nsa1-Rrp1-Rpf1-Mak16模块稳定了60S亚基的溶剂侧，而Erb1- ytm1 - nop7复合体通过Erb1的弯曲n端延伸、8个组装因子、3个核糖体蛋白和3个25S rRNA结构域进行组织和连接。我们的结构快照揭示了早期核仁60S颗粒中六个主要60S结构域的整合和压实顺序，这些结构域从溶剂侧沿着出口隧道逐步发展到中央凸起。cro - em组装因子Nsa1、Mak16、Rpf1和Rrp1在溶剂侧构成一个模块，Erb1在亚基间侧充当中心协调者，在pre-rRNA的5 ‘到3 ’圆形成后，有一个顺序组装途径。cro - em对pre-60S核糖体结构的分析提供了对核糖体组装至关重要的顺序事件和中间状态以及许多相关因子的功能的见解。
Eukaryotic 60S ribosomal subunits are comprised of three rRNAs and ∼50 ribosomal proteins. The initial steps of their formation take place in the nucleolus, but, owing to a lack of structural information, this process is poorly understood. Using cryo-EM, we solved structures of early 60S biogenesis intermediates at 3.3 Å to 4.5 Å resolution, thereby providing insights into their sequential folding and assembly pathway. Besides revealing distinct immature rRNA conformations, we map 25 assembly factors in six different assembly states. Notably, the Nsa1-Rrp1-Rpf1-Mak16 module stabilizes the solvent side of the 60S subunit, and the Erb1-Ytm1-Nop7 complex organizes and connects through Erb1’s meandering N-terminal extension, eight assembly factors, three ribosomal proteins, and three 25S rRNA domains. Our structural snapshots reveal the order of integration and compaction of the six major 60S domains within early nucleolar 60S particles developing stepwise from the solvent side around the exit tunnel to the central protuberance. Structures of five nucleolar pre-60S intermediates by cryo-EM Assembly factors Nsa1, Mak16, Rpf1, and Rrp1 form a module at the solvent side Erb1 acts as a central coordinator at the intersubunit side A sequential assembly pathway follows after the 5′ to 3′ circular formation of pre-rRNA Cryo-EM analysis of the architecture of pre-60S ribosomes provides insights into the sequential events and intermediate states critical for ribosome assembly, as well as the functions of many associated factors.
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影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ
影响因子：
10.5
作者：
Gamalinda M;Ohmayer U;Jakovljevic J;Kumcuoglu B;Woolford J;Mbom B;Lin L;Woolford JL Jr
通讯作者：
Woolford JL Jr
影响因子：
16.8
作者：
Barrio-Garcia, Clara;Thoms, Matthias;Hurt, Ed
通讯作者：
Hurt, Ed
影响因子：
16
作者：
Grandi, P;Rybin, V;Hurt, E
通讯作者：
Hurt, E
DOI：
10.1107/s0907444909042073
发表时间：
2010-01
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Chen VB;Arendall WB 3rd;Headd JJ;Keedy DA;Immormino RM;Kapral GJ;Murray LW;Richardson JS;Richardson DC
通讯作者：
Richardson DC