1Sahlgrenska Cancer Center, Institute of Biomedicine, Department of Pathology, Sahlgrenska Academy, University of Gothenburg, Box 425, 40530, Gothenburg, Sweden.
FASEB J. 2013 Dec;27(12):4965-74. doi: 10.1096/fj.13-234435. Epub 2013 Aug 23.
The three FET (FUS, EWSR1, and TAF15) family RNA binding proteins are expressed in all tissues and almost all cell types. The disordered N-terminal parts are always present in FET fusion oncoproteins of sarcomas and leukemia. Mutations in FUS and TAF15 cause aggregation of FET proteins in neurological disorders. Here we used recombinant proteins in pulldown experiments and mass spectrometry to identify major interaction partners of the FET N-terminal parts. We report that FUS, EWSR1, and TAF15 form homo- and heterocomplexes as major binding partners and identify an evolutionarily conserved N-terminal motif (FETBM1) that is required for this interaction. The binding is RNA and DNA independent and robust up to 1 M of NaCl. The localization of FETBM1 and its target sequences supports a simple model for FET protein aggregation as reported in neurological disorders such as amyotrophic lateral sclerosis, frontotemporal dementia, and essential tremor. The FETBM1 localization also explains the binding of normal full-length FET proteins to their oncogenic fusion proteins.
三种 FET(FUS、EWSR1 和 TAF15)家族 RNA 结合蛋白在所有组织和几乎所有细胞类型中均有表达。在肉瘤和白血病的 FET 融合癌蛋白中,无序的 N 端部分总是存在。FUS 和 TAF15 的突变导致 FET 蛋白在神经退行性疾病中聚集。在这里,我们使用重组蛋白在下拉实验和质谱中鉴定 FET N 端部分的主要相互作用伙伴。我们报告 FUS、EWSR1 和 TAF15 形成同源和异源复合物作为主要结合伙伴,并鉴定出一个进化上保守的 N 端基序(FETBM1),该基序对于这种相互作用是必需的。该结合不依赖于 RNA 和 DNA,并且在高达 1 M 的 NaCl 下仍然稳定。FETBM1 的定位及其靶序列支持神经退行性疾病(如肌萎缩性侧索硬化症、额颞叶痴呆和特发性震颤)中报告的 FET 蛋白聚集的简单模型。FETBM1 的定位也解释了正常全长 FET 蛋白与其致癌融合蛋白的结合。