Detection of 3'-end RNA uridylation with a protein nanopore.

Detection of 3'-end RNA uridylation with a protein nanopore.
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DOI:
10.1021/nn4050479
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发表时间:
2014-02-25
期刊:
影响因子:
17.1
通讯作者:
Bayley H
Bayley H
中科院分区:
材料科学1区
文献类型:
--
作者:
Clamer M;Höfler L;Mikhailova E;Viero G;Bayley H

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RNA分子3′-末端的转录后修饰对其在细胞中的稳定性和加工有着深远的影响。最近发现,尿苷化,即在3′-末端添加尿苷,是稳定标记分子或指导其降解的重要调节信号。用于检测这种转录后修饰的简单且具有成本效益的方法尚不可用。在这里,我们证明了3′-尿苷化的ssRNA在葡萄球菌α-溶血素(αHL)纳米孔的β桶内的选择性和瞬时结合,并研究了孔识别尿苷的分子基础。我们显示了基于其长度的3′-寡尿苷尾的区分,并提出αHL纳米孔作为这种生物相关RNA修饰的有用传感器。
Post-transcriptional modifications of the 3′-ends of RNA molecules have a profound impact on their stability and processing in the cell. Uridylation, the addition of uridines to 3′-ends, has recently been found to be an important regulatory signal to stabilize the tagged molecules or to direct them towards degradation. Simple and cost-effective methods for the detection of this post-transcriptional modification are not yet available. Here, we demonstrate the selective and transient binding of 3′-uridylated ssRNAs inside the β barrel of the staphylococcal alpha-hemolysin (αHL) nanopore, and investigate the molecular basis of uridine recognition by the pore. We show the discrimination of 3′-oligouridine tails on the basis of their lengths and propose the αHL nanopore as a useful sensor for this biologically relevant RNA modification.
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