Jin S-H, Yao Y, Wu Q
МОЕ Key Lab. Bioinformatics, Tsinghua University, Beijing, 100084 China.
Center for Synthetic and Systems Biology, Tsinghua University, Beijing, 100084 China.
Mol Biol (Mosk). 2019 Jan-Feb;53(1):64-73. doi: 10.1134/S002689841901004X.
The previously developed technology RNA enhancement (RNAe) is reported to increase specific gene expression at post-transcriptional level via a long noncoding RNA (lncRNA). The mechanism for SINEB2-dependent enhancement of translation remains not well understood. Here we present the result of experiments with the folded states of IncRNA in doubly deionized water obtained by slowcool method. These IncRNA were used in RNA pull-down assay that yielded six IncRNA-binding proteins potentially involved in RNAe. The direct interactions of IncRNA with interleukin enhancer-binding factor 3 (ILF3) and eukaryotic initiation factor 4A-I (eIF4Al) in vivo and in vitro were confirmed in RNA-binding protein affinity experiment and electrophoretic mobility shift assay (EMSA), respectively. These observations could explain RNAe phenomenon through IncRNA-dependent guiding of ILF3 protein, which, in turn, recruits polysomes or the factors for translation initiation, and attracting eIF4Al proteins accelerating the unwinding of the secondary structure at the 5'-end of mRNA during translation initiation. Therefore, the hypothetical mechanism through which IncRNAs may regulate the translation of a specific mRNA is proposed.
据报道,先前开发的技术RNA增强(RNAe)可通过长链非编码RNA(lncRNA)在转录后水平增加特定基因的表达。SINEB2依赖性翻译增强的机制仍未完全了解。在这里,我们展示了通过慢冷法在双去离子水中获得的lncRNA折叠状态的实验结果。这些lncRNA用于RNA下拉实验,该实验产生了六种可能参与RNAe的lncRNA结合蛋白。在RNA结合蛋白亲和力实验和电泳迁移率变动分析(EMSA)中,分别证实了lncRNA与白细胞介素增强子结合因子3(ILF3)和真核起始因子4A-I(eIF4A1)在体内和体外的直接相互作用。这些观察结果可以通过lncRNA依赖性指导ILF3蛋白来解释RNAe现象,ILF3蛋白进而招募多核糖体或翻译起始因子,并吸引eIF4A1蛋白在翻译起始过程中加速mRNA 5'端二级结构的解旋。因此,提出了lncRNA可能调节特定mRNA翻译的假设机制。