抑制 RNA 剪接会触发 CHMP7 进入细胞核,影响 TDP-43 的功能,导致 ALS 细胞表型的出现。

IF 14.7 1区 医学 Q1 NEUROSCIENCES
Neuron Pub Date : 2024-12-18 Epub Date: 2024-10-31 DOI:10.1016/j.neuron.2024.10.007
Norah Al-Azzam, Jenny H To, Vaishali Gautam, Lena A Street, Chloe B Nguyen, Jack T Naritomi, Dylan C Lam, Assael A Madrigal, Benjamin Lee, Wenhao Jin, Anthony Avina, Orel Mizrahi, Jasmine R Mueller, Willard Ford, Cara R Schiavon, Elena Rebollo, Anthony Q Vu, Steven M Blue, Yashwin L Madakamutil, Uri Manor, Jeffrey D Rothstein, Alyssa N Coyne, Marko Jovanovic, Gene W Yeo
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引用次数: 0

摘要

肌萎缩性脊髓侧索硬化症(ALS)与神经元中某些核蛋白的减少有关。带电多囊体蛋白 7(CHMP7)是一种参与核孔监控的蛋白质,它的核定位增加已被确定为破坏核孔和干扰运输的一个关键因素。利用基于 CRISPR 的 microRaft 和 gRNA 鉴定(CRaft-ID),我们发现了 55 个影响 CHMP7 定位的 RNA 结合蛋白(RBPs),其中包括运动神经元(SMN)生存复合体成分 SmD1。免疫沉淀-质谱(IP-MS)和增强交联与免疫沉淀(CLIP)分析揭示了CHMP7与运动神经元(MNs)中的SmD1、小核RNA和剪接因子mRNA的相互作用。ALS 诱导多能干细胞(iPSC)-MNs 显示 SmD1 表达减少,抑制 SmD1/SMN 复合物可增加 CHMP7 的核定位。重要的是,在ALS iPSC-MNs中过表达SmD1可恢复CHMP7的胞浆定位并纠正STMN2的剪接。我们的研究结果表明,ALS 早期发病机制是由 SMN 复合物失调驱动的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Inhibition of RNA splicing triggers CHMP7 nuclear entry, impacting TDP-43 function and leading to the onset of ALS cellular phenotypes.

Amyotrophic lateral sclerosis (ALS) is linked to the reduction of certain nucleoporins in neurons. Increased nuclear localization of charged multivesicular body protein 7 (CHMP7), a protein involved in nuclear pore surveillance, has been identified as a key factor damaging nuclear pores and disrupting transport. Using CRISPR-based microRaft, followed by gRNA identification (CRaft-ID), we discovered 55 RNA-binding proteins (RBPs) that influence CHMP7 localization, including SmD1, a survival of motor neuron (SMN) complex component. Immunoprecipitation-mass spectrometry (IP-MS) and enhanced crosslinking and immunoprecipitation (CLIP) analyses revealed CHMP7's interactions with SmD1, small nuclear RNAs, and splicing factor mRNAs in motor neurons (MNs). ALS induced pluripotent stem cell (iPSC)-MNs show reduced SmD1 expression, and inhibiting SmD1/SMN complex increased CHMP7 nuclear localization. Crucially, overexpressing SmD1 in ALS iPSC-MNs restored CHMP7's cytoplasmic localization and corrected STMN2 splicing. Our findings suggest that early ALS pathogenesis is driven by SMN complex dysregulation.

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来源期刊
Neuron
Neuron 医学-神经科学
CiteScore
24.50
自引率
3.10%
发文量
382
审稿时长
1 months
期刊介绍: Established as a highly influential journal in neuroscience, Neuron is widely relied upon in the field. The editors adopt interdisciplinary strategies, integrating biophysical, cellular, developmental, and molecular approaches alongside a systems approach to sensory, motor, and higher-order cognitive functions. Serving as a premier intellectual forum, Neuron holds a prominent position in the entire neuroscience community.
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