In Vivo RNAi-Mediated eIF3m Knockdown Affects Ribosome Biogenesis and Transcription but Has Limited Impact on mRNA-Specific Translation.
In Vivo RNAi-Mediated eIF3m Knockdown Affects Ribosome Biogenesis and Transcription but Has Limited Impact on mRNA-Specific Translation.
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体内 RNAi 介导的 eIF3m 敲低会影响核糖体生物合成和转录，但对 mRNA 特异性翻译的影响有限。
DOI：
10.1016/j.omtn.2019.11.009
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Koteliansky,Vict
中科院分区：
文献类型：
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作者：
Smekalova,ElenaM;Gerashchenko,MaximV;O'Connor,PatrickBF;Whittaker,CharlesA;Kauffman,KevinJ;Fefilova,AnnaS;Zatsepin,TimofeiS;Bogorad,RomanL;Baranov,PavelV;Langer,Robert;Gladyshev,VadimN;Anderson,DanielG;Koteliansky,Vict
翻译是一个重要的生物学过程，失调与一系列疾病有关，包括核糖体病、糖尿病和癌症。在这里，我们研究翻译失调在vivousing RNAi敲低小鼠肝脏中的翻译起始因子eIF3的m亚基。转录组测序、核糖体分析、全蛋白质组和磷酸化蛋白质组分析表明，eIF3m缺陷导致转录反应和细胞翻译的变化，在特定mRNA的翻译中产生很少可检测的差异。转录反应分为两大类：核糖体生物发生（核糖体蛋白质转录增加）和细胞代谢（脂质、氨基酸、核酸和药物代谢的改变）。核糖体生物发生的分析揭示了rRNA加工的抑制，突出了响应eIF3m敲低的rRNA合成和核糖体蛋白基因转录的解耦。有趣的是，eIF3m蛋白水平的类似降低与体外mTOR通路的诱导有关，但与体内无关。总之，这项工作突出了利用RNAi为基础的体内方法研究哺乳动物体内的调节。
Translation is an essential biological process, and dysregulation is associated with a range of diseases including ribosomopathies, diabetes, and cancer. Here, we examine translation dysregulationin vivousing RNAi to knock down the m-subunit of the translation initiation factor eIF3 in the mouse liver. Transcriptome sequencing, ribosome profiling, whole proteome, and phosphoproteome analyses show that eIF3m deficiency leads to the transcriptional response and changes in cellular translation that yield few detectable differences in the translation of particular mRNAs. The transcriptional response fell into two main categories: ribosome biogenesis (increased transcription of ribosomal proteins) and cell metabolism (alterations in lipid, amino acid, nucleic acid, and drug metabolism). Analysis of ribosome biogenesis reveals inhibition of rRNA processing, highlighting decoupling of rRNA synthesis and ribosomal protein gene transcription in response to eIF3m knockdown. Interestingly, a similar reduction in eIF3m protein levels is associated with induction of the mTOR pathwayin vitrobut notin vivo. Overall, this work highlights the utility of a RNAi-basedin vivoapproach for studying the regulation of mammalian translationin vivo.