Laboratory of Molecular Neuro-Oncology, The Rockefeller University, New York, NY 10065, USA.
Cell. 2011 Jul 22;146(2):247-61. doi: 10.1016/j.cell.2011.06.013.
FMRP loss of function causes Fragile X syndrome (FXS) and autistic features. FMRP is a polyribosome-associated neuronal RNA-binding protein, suggesting that it plays a key role in regulating neuronal translation, but there has been little consensus regarding either its RNA targets or mechanism of action. Here, we use high-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation (HITS-CLIP) to identify FMRP interactions with mouse brain polyribosomal mRNAs. FMRP interacts with the coding region of transcripts encoding pre- and postsynaptic proteins and transcripts implicated in autism spectrum disorders (ASD). We developed a brain polyribosome-programmed translation system, revealing that FMRP reversibly stalls ribosomes specifically on its target mRNAs. Our results suggest that loss of a translational brake on the synthesis of a subset of synaptic proteins contributes to FXS. In addition, they provide insight into the molecular basis of the cognitive and allied defects in FXS and ASD and suggest multiple targets for clinical intervention.
脆性 X 综合征（FXS）和自闭症特征是由 FMRP 功能丧失引起的。FMRP 是一种多核糖体相关的神经元 RNA 结合蛋白，这表明它在调节神经元翻译中发挥着关键作用，但关于其 RNA 靶标或作用机制尚未达成共识。在这里，我们使用交联免疫沉淀（HITS-CLIP）分离的 RNA 的高通量测序来鉴定 FMRP 与小鼠脑多核糖体 mRNA 的相互作用。FMRP 与编码前突触和后突触蛋白以及与自闭症谱系障碍（ASD）相关的转录本的编码区相互作用。我们开发了一种脑多核糖体程序化翻译系统，揭示 FMRP 可特异性地可逆地使核糖体停滞在其靶 mRNA 上。我们的研究结果表明，合成一组特定的突触蛋白的翻译被阻断，这可能导致 FXS。此外，它们为 FXS 和 ASD 的认知和相关缺陷的分子基础提供了线索，并为临床干预提供了多个靶点。
