FUS-RNA结合结构域和低复杂性结构域在RNA依赖性相分离中的作用。
The roles of FUS-RNA binding domain and low complexity domain in RNA-dependent phase separation.
作者信息
Ganser Laura R, Niaki Amirhossein Ghanbari, Yuan Xincheng, Huang Ethan, Deng Dahlia, Djaja Nathalie A, Ge Yingda, Craig Alanna, Langlois Olivia, Myong Sua
机构信息
Department of Biophysics, Johns Hopkins University, Baltimore, MD 21218, USA.
Department of Biophysics, Johns Hopkins University, Baltimore, MD 21218, USA; Program in Cell, Molecular, Developmental Biology, and Biophysics, Johns Hopkins University, 3400 N Charles St, Baltimore, MD 21218, USA; Department of Biology, Johns Hopkins University, 3400 N Charles St, Baltimore, MD 21218, USA.
出版信息
Structure. 2024 Feb 1;32(2):177-187.e5. doi: 10.1016/j.str.2023.11.006. Epub 2023 Dec 8.
Fused in sarcoma (FUS) is an archetypal phase separating protein asymmetrically divided into a low complexity domain (LCD) and an RNA binding domain (RBD). Here, we explore how the two domains contribute to RNA-dependent phase separation, RNA recognition, and multivalent complex formation. We find that RBD drives RNA-dependent phase separation but forms large and irregularly shaped droplets that are rescued by LCD in trans. Electrophoretic mobility shift assay (EMSA) and single-molecule fluorescence assays reveal that, while both LCD and RBD bind RNA, RBD drives RNA engagement and multivalent complex formation. While RBD alone exhibits delayed RNA recognition and a less dynamic RNP complex compared to full-length FUS, LCD in trans rescues full-length FUS activity. Likewise, cell-based data show RBD forms nucleolar condensates while LCD in trans rescues the diffuse nucleoplasm localization of full-length FUS. Our results point to a regulatory role of LCD in tuning the RNP interaction and buffering phase separation.
肉瘤融合蛋白（FUS）是一种典型的相分离蛋白，不对称地分为低复杂性结构域（LCD）和RNA结合结构域（RBD）。在此，我们探究这两个结构域如何促进RNA依赖性相分离、RNA识别以及多价复合物形成。我们发现RBD驱动RNA依赖性相分离，但形成的液滴大且形状不规则，而LCD可通过反式作用挽救这种情况。电泳迁移率变动分析（EMSA）和单分子荧光分析表明，虽然LCD和RBD都能结合RNA，但RBD驱动RNA结合和多价复合物形成。与全长FUS相比，单独的RBD表现出延迟的RNA识别和动态性较低的核糖核蛋白复合物，而LCD的反式作用可挽救全长FUS的活性。同样，基于细胞的数据表明，RBD形成核仁凝聚物，而LCD的反式作用可挽救全长FUS在核质中的弥散定位。我们的结果表明，LCD在调节核糖核蛋白相互作用和缓冲相分离方面具有调节作用。
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