Department of Molecular Genetics and Microbiology, Duke University Medical Center, Durham, NC 27710, USA.
Mol Cell. 2011 Aug 5;43(3):327-39. doi: 10.1016/j.molcel.2011.06.007. Epub 2011 Jun 30.
RNA-binding proteins coordinate the fates of multiple RNAs, but the principles underlying these global interactions remain poorly understood. We elucidated regulatory mechanisms of the RNA-binding protein HuR, by integrating data from diverse high-throughput targeting technologies, specifically PAR-CLIP, RIP-chip, and whole-transcript expression profiling. The number of binding sites per transcript, degree of HuR association, and degree of HuR-dependent RNA stabilization were positively correlated. Pre-mRNA and mature mRNA containing both intronic and 3' UTR binding sites were more highly stabilized than transcripts with only 3' UTR or only intronic binding sites, suggesting that HuR couples pre-mRNA processing with mature mRNA stability. We also observed HuR-dependent splicing changes and substantial binding of HuR in polypyrimidine tracts of pre-mRNAs. Comparison of the spatial patterns surrounding HuR and miRNA binding sites provided functional evidence for HuR-dependent antagonism of proximal miRNA-mediated repression. We conclude that HuR coordinates gene expression outcomes at multiple interconnected steps of RNA processing.
RNA 结合蛋白协调多种 RNA 的命运，但这些全局相互作用的原理仍知之甚少。我们通过整合来自多种高通量靶向技术的数据，特别是 PAR-CLIP、RIP-chip 和全转录表达谱分析，阐明了 RNA 结合蛋白 HuR 的调控机制。每个转录本的结合位点数量、HuR 结合程度和 HuR 依赖性 RNA 稳定性程度呈正相关。含有内含子和 3'UTR 结合位点的 pre-mRNA 和成熟 mRNA 比仅含有 3'UTR 或仅含有内含子结合位点的转录本更稳定，这表明 HuR 将 pre-mRNA 加工与成熟 mRNA 稳定性联系起来。我们还观察到 HuR 依赖性剪接变化和 HuR 在 pre-mRNA 的多嘧啶区的大量结合。HuR 结合位点和 miRNA 结合位点周围空间模式的比较为 HuR 依赖性拮抗近端 miRNA 介导的抑制提供了功能证据。我们得出结论，HuR 在 RNA 加工的多个相互关联的步骤中协调基因表达结果。
