Biomolecular condensation of NUP98 fusion proteins drives leukemogenic gene expression
Biomolecular condensation of NUP98 fusion proteins drives leukemogenic gene expression
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DOI：
10.1038/s41594-020-00550-w
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发表时间：
2021-01-21
影响因子：
16.8
通讯作者：
Grebien, Florian
中科院分区：
文献类型：
--
作者：
Terlecki-Zaniewicz, Stefan;Humer, Theresa;Grebien, Florian
利用质谱技术研究了不同NUP 98融合蛋白引起白血病的机制，发现NUP 98融合蛋白具有共同的相互作用因子，改变了核凝聚物的组成，NUP 98融合蛋白通过未知的分子机制引起白血病。所有NUP 98融合蛋白在NUP 98 N末端共有一个固有无序区（IDR），其特征在于苯丙氨酸-甘氨酸（FG）的重复，并且C末端融合伴侣通常在基因控制中起作用。我们研究了NUP 98融合蛋白的致癌转化机制是否与其蛋白质相互作用组有关。亲和纯化耦合到质谱（MS）和共聚焦成像的五个NUP 98融合蛋白在人白血病细胞中表达的揭示，共享的相互作用富集的蛋白质参与生物分子凝聚，他们共定位与NUP 98融合蛋白在核斑点。我们开发了生物素化异恶唑介导的凝聚体MS（biCon-MS），以显示NUP 98融合蛋白改变生物分子凝聚物的整体组成。一种含有FG重复序列的人工融合蛋白表型模仿NUP 98融合蛋白的核定位模式及其驱动致癌基因表达程序的能力。因此，我们提出含IDR的融合蛋白联合收割机结合生物分子凝聚与转录控制来诱导癌症。
A mass spectrometry-based approach is used to investigate the mechanisms by which different NUP98 fusion proteins cause leukemia, revealing that the fusion proteins share common interactors and alter the composition of nuclear condensates.NUP98 fusion proteins cause leukemia via unknown molecular mechanisms. All NUP98 fusion proteins share an intrinsically disordered region (IDR) in the NUP98 N terminus, featuring repeats of phenylalanine-glycine (FG), and C-terminal fusion partners often function in gene control. We investigated whether mechanisms of oncogenic transformation by NUP98 fusion proteins are hardwired in their protein interactomes. Affinity purification coupled to mass spectrometry (MS) and confocal imaging of five NUP98 fusion proteins expressed in human leukemia cells revealed that shared interactors were enriched for proteins involved in biomolecular condensation and that they colocalized with NUP98 fusion proteins in nuclear puncta. We developed biotinylated isoxazole-mediated condensome MS (biCon-MS) to show that NUP98 fusion proteins alter the global composition of biomolecular condensates. An artificial FG-repeat-containing fusion protein phenocopied the nuclear localization patterns of NUP98 fusion proteins and their capability to drive oncogenic gene expression programs. Thus, we propose that IDR-containing fusion proteins combine biomolecular condensation with transcriptional control to induce cancer.