Rrp5 binding at multiple sites coordinates pre-rRNA processing and assembly.
Rrp5 binding at multiple sites coordinates pre-rRNA processing and assembly.
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DOI：
10.1016/j.molcel.2013.10.017
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发表时间：
2013-12-12
期刊：
影响因子：
16
通讯作者：
Tollervey, David
中科院分区：
文献类型：
--
作者：
Lebaron, Simon;Segerstolpe, Asa;French, Sarah L.;Dudnakova, Tatiana;Alves, Flavia de Lima;Granneman, Sander;Rappsilber, Juri;Beyer, Ann L.;Wieslander, Lars;Tollervey, David
体内紫外线交联鉴定出大量高度保守的核糖体合成因子 Rrp5 的前核糖体 RNA (pre-rRNA) 结合位点。分子内互补表明，Rrp5 的 C 端结构域 (CTD) 是 18S rRNA 合成途径中 A0-A2 位点前 rRNA 裂解所需的，而 N 端结构域 (NTD) 是 5.8S/25S rRNA 合成途径中 A3 裂解所需的。 CTD 与 A2 侧翼序列以及 snoRNA U3、U14、snR30 和 snR10 交联，这些是 A0-A2 处切割所需的。 NTD 与 A3 侧翼序列和核糖核酸酶 MRP 的 RNA 成分交联，后者切割 A3 位点。 Rrp5 还可以直接与几种大结构蛋白和核苷三磷酸酶交联。染色质扩散证实了协调前核糖体组装和加工的关键作用。 Rrp5 耗尽后，共转录切割消失，前核糖体压缩大大减少。 Rrp5 结合前 rRNA 中的多个分散位点 Rrp5 的 NTD 和 CTD 各自结合到需要它们的切割位点附近 Rrp5 直接结合大的结构蛋白和 NTPase Rrp5 是前核糖体压缩所必需的
In vivo UV crosslinking identified numerous preribosomal RNA (pre-rRNA) binding sites for the large, highly conserved ribosome synthesis factor Rrp5. Intramolecular complementation has shown that the C-terminal domain (CTD) of Rrp5 is required for pre-rRNA cleavage at sites A0–A2 on the pathway of 18S rRNA synthesis, whereas the N-terminal domain (NTD) is required for A3 cleavage on the pathway of 5.8S/25S rRNA synthesis. The CTD was crosslinked to sequences flanking A2 and to the snoRNAs U3, U14, snR30, and snR10, which are required for cleavage at A0–A2. The NTD was crosslinked to sequences flanking A3 and to the RNA component of ribonuclease MRP, which cleaves site A3. Rrp5 could also be directly crosslinked to several large structural proteins and nucleoside triphosphatases. A key role in coordinating preribosomal assembly and processing was confirmed by chromatin spreads. Following depletion of Rrp5, cotranscriptional cleavage was lost and preribosome compaction greatly reduced. Rrp5 binds multiple dispersed sites in the pre-rRNA The NTD and CTD of Rrp5 each bind adjacent to sites of cleavages that require them Rrp5 directly binds large, structural proteins and NTPases Rrp5 is required for preribosome compaction
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