Ribosomal protein L35 is required for 27SB pre-rRNA processing in Saccharomyces cerevisiae.

Ribosomal protein L35 is required for 27SB pre-rRNA processing in Saccharomyces cerevisiae.
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DOI:
10.1093/nar/gkq260
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发表时间:
2010-08
影响因子:
14.9
通讯作者:
de la Cruz J
de la Cruz J
中科院分区:
生物学2区
文献类型:
--
作者:
Babiano R;de la Cruz J

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核糖体的合成涉及前rRNA加工和核糖体蛋白组装的伴随。在真核生物中,这是一个复杂的过程,需要前体rRNA内的特定序列和结构、至少200个反式作用因子和核糖体蛋白的参与。关于单个60 S核糖体蛋白在核糖体合成中的功能的信息很少。在此,我们分析了核糖体蛋白L35在核糖体生物发生中的作用。体内L35缺失导致60 S核糖体亚基缺陷和半聚体多聚体的出现。脉冲追踪、北方杂交和引物延伸分析显示,27 SB至7 S前体rRNA的加工在L35耗尽后强烈延迟。最可能的结果是,前60 S核糖体颗粒从核仁释放到核质也被阻断。RPL 35 A的缺失导致相似但不太明显的表型。此外,我们表明,L35组装在核仁中,并结合到早期前60 S核糖体颗粒。最后,流式细胞术分析表明,L35-耗尽的细胞轻度延迟细胞周期的G1期。我们的结论是,L35的组装是一个先决条件的有效切割的内部转录间隔区2在网站C2。
Ribosome synthesis involves the concomitance of pre-rRNA processing and ribosomal protein assembly. In eukaryotes, this is a complex process that requires the participation of specific sequences and structures within the pre-rRNAs, at least 200 trans-acting factors and the ribosomal proteins. There is little information on the function of individual 60S ribosomal proteins in ribosome synthesis. Herein, we have analysed the contribution of ribosomal protein L35 in ribosome biogenesis. In vivo depletion of L35 results in a deficit in 60S ribosomal subunits and the appearance of half-mer polysomes. Pulse-chase, northern hybridization and primer extension analyses show that processing of the 27SB to 7S pre-rRNAs is strongly delayed upon L35 depletion. Most likely as a consequence of this, release of pre-60S ribosomal particles from the nucleolus to the nucleoplasm is also blocked. Deletion of RPL35A leads to similar although less pronounced phenotypes. Moreover, we show that L35 assembles in the nucleolus and binds to early pre-60S ribosomal particles. Finally, flow cytometry analysis indicated that L35-depleted cells mildly delay the G1 phase of the cell cycle. We conclude that L35 assembly is a prerequisite for the efficient cleavage of the internal transcribed spacer 2 at site C2.
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