Max-Delbrück-Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 13125 Berlin, Germany.
Max-Delbrück-Center for Molecular Medicine, Berlin Institute for Medical Systems Biology, 13125 Berlin, Germany.
Mol Cell. 2014 May 22;54(4):573-85. doi: 10.1016/j.molcel.2014.03.017. Epub 2014 Apr 10.
RNA helicases are important regulators of gene expression that act by remodeling RNA secondary structures and RNA-protein interactions. Here, we demonstrate that MOV10 has an ATP-dependent 5' to 3' in vitro RNA unwinding activity and determine the RNA-binding sites of MOV10 and its helicase mutants using PAR-CLIP. We find that MOV10 predominantly binds to 3' UTRs upstream of regions predicted to form local secondary structures and provide evidence that MOV10 helicase mutants are impaired in their ability to translocate 5' to 3' on their mRNA targets. MOV10 interacts with UPF1, the key component of the nonsense-mediated mRNA decay pathway. PAR-CLIP of UPF1 reveals that MOV10 and UPF1 bind to RNA in close proximity. Knockdown of MOV10 resulted in increased mRNA half-lives of MOV10-bound as well as UPF1-regulated transcripts, suggesting that MOV10 functions in UPF1-mediated mRNA degradation as an RNA clearance factor to resolve structures and displace proteins from 3' UTRs.
RNA 解旋酶是重要的基因表达调控因子，通过重塑 RNA 二级结构和 RNA-蛋白质相互作用来发挥作用。在这里，我们证明 MOV10 具有 ATP 依赖性的体外 5' 到 3' RNA 解链活性，并使用 PAR-CLIP 确定了 MOV10 及其解旋酶突变体的 RNA 结合位点。我们发现 MOV10 主要结合到预测形成局部二级结构的区域上游的 3'UTR，并提供证据表明 MOV10 解旋酶突变体在其 mRNA 靶标上从 5' 到 3' 易位的能力受损。MOV10 与 UPF1 相互作用，UPF1 是无意义介导的 mRNA 降解途径的关键组成部分。UPF1 的 PAR-CLIP 揭示了 MOV10 和 UPF1 紧密结合在 RNA 上。MOV10 的敲低导致与 MOV10 结合的以及 UPF1 调节的转录本的 mRNA 半衰期增加，表明 MOV10 作为 RNA 清除因子在 UPF1 介导的 mRNA 降解中发挥作用，以解决结构并将蛋白质从 3'UTR 中置换。
