Department of Pathology and Cell Biology, Institute for Research in Immunology and Cancer, University of Montreal, Montreal, QC, Canada.
Institute for Research in Immunology and Cancer (IRIC), Université de Montréal, Montreal, QC, Canada.
EMBO J. 2023 Apr 3;42(7):e110496. doi: 10.15252/embj.2021110496. Epub 2023 Feb 27.
Aberrant splicing is typically attributed to splice-factor (SF) mutation and contributes to malignancies including acute myeloid leukemia (AML). Here, we discovered a mutation-independent means to extensively reprogram alternative splicing (AS). We showed that the dysregulated expression of eukaryotic translation initiation factor eIF4E elevated selective splice-factor production, thereby impacting multiple spliceosome complexes, including factors mutated in AML such as SF3B1 and U2AF1. These changes generated a splicing landscape that predominantly supported altered splice-site selection for ~800 transcripts in cell lines and ~4,600 transcripts in specimens from high-eIF4E AML patients otherwise harboring no known SF mutations. Nuclear RNA immunoprecipitations, export assays, polysome analyses, and mutational studies together revealed that eIF4E primarily increased SF production via its nuclear RNA export activity. By contrast, eIF4E dysregulation did not induce known SF mutations or alter spliceosome number. eIF4E interacted with the spliceosome and some pre-mRNAs, suggesting its direct involvement in specific splicing events. eIF4E induced simultaneous effects on numerous SF proteins, resulting in a much larger range of splicing alterations than in the case of mutation or dysregulation of individual SFs and providing a novel paradigm for splicing control and dysregulation.
异常剪接通常归因于剪接因子 (SF) 突变,并导致包括急性髓细胞性白血病 (AML) 在内的恶性肿瘤。在这里,我们发现了一种非突变依赖性的广泛重编程选择性剪接 (AS) 的方法。我们表明,真核翻译起始因子 eIF4E 的失调表达会升高选择性剪接因子的产生,从而影响多个剪接体复合物,包括在 AML 中发生突变的因子,如 SF3B1 和 U2AF1。这些变化产生了一种剪接图谱,主要支持约 800 个细胞系中的转录本和约 4600 个高 eIF4E AML 患者标本中的转录本的改变剪接位点选择,而这些患者没有已知的 SF 突变。核 RNA 免疫沉淀、出口测定、多核糖体分析和突变研究共同揭示,eIF4E 主要通过其核 RNA 输出活性增加 SF 产生。相比之下,eIF4E 失调不会诱导已知的 SF 突变或改变剪接体数量。eIF4E 与剪接体和一些前体 mRNA 相互作用,表明其直接参与特定的剪接事件。eIF4E 对许多 SF 蛋白同时产生影响,导致比单个 SF 突变或失调的情况下更大范围的剪接改变,并为剪接控制和失调提供了一个新的范例。