Fusion protein EWS-FLI1 is incorporated into a protein granule in cells.
Fusion protein EWS-FLI1 is incorporated into a protein granule in cells.
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DOI：
10.1261/rna.078827.121
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发表时间：
2021-05-25
期刊：
影响因子：
--
通讯作者：
Schwartz JC
中科院分区：
文献类型：
--
作者：
Ahmed NS;Harrell LM;Wieland DR;Lay MA;Thompson VF;Schwartz JC
尤文氏肉瘤是由含有低复杂性结构域的融合蛋白驱动的，该结构域本质上是无序的，是一种强大的转录调节因子。EWS-FLI1是在Ewing肉瘤中发现的最常见的融合蛋白，其LC结构域来自rna结合蛋白EWSR1 (Ewing肉瘤rna结合蛋白1)，dna结合结构域来自转录因子FLI1 （Friend白血病病毒整合1）。EWS-FLI1可以结合RNA聚合酶II （RNA Pol II）并通过LC结构域进行自组装。rna结合蛋白如EWSR1在细胞中自组装或相分离的能力引起了人们对这一过程对EWS-FLI1活性的贡献的质疑。我们通过sirna介导的敲低和RNA-seq分析检测了Ewing肉瘤细胞中EWSR1和EWS-FLI1的活性。受EWSR1敲低影响的转录本比预期的要多，其中包括许多EWS-FLI1调控基因。我们重新评估了EWS-FLI1、EWSR1和RNA Pol II之间的物理相互作用，并使用基于交联的策略来研究与蛋白质相关的蛋白质组装。EWS-FLI1的LC结构域是观察到的组装在细胞中形成所必需的。这些结果为蛋白质组装提供了新的见解，该蛋白质组装可能使EWS-FLI1结合其广泛的蛋白质伴侣网络，并有助于调节Ewing肉瘤的基因表达。
Ewing sarcoma is driven by fusion proteins containing a low-complexity (LC) domain that is intrinsically disordered and a powerful transcriptional regulator. The most common fusion protein found in Ewing sarcoma, EWS-FLI1, takes its LC domain from the RNA-binding protein EWSR1 (Ewing sarcoma RNA-binding protein 1) and a DNA-binding domain from the transcription factor FLI1 (Friend leukemia virus integration 1). EWS-FLI1 can bind RNA polymerase II (RNA Pol II) and self-assemble through its LC domain. The ability of RNA-binding proteins like EWSR1 to self-assemble or phase separate in cells has raised questions about the contribution of this process to EWS-FLI1 activity. We examined EWSR1 and EWS-FLI1 activity in Ewing sarcoma cells by siRNA-mediated knockdown and RNA-seq analysis. More transcripts were affected by the EWSR1 knockdown than expected and these included many EWS-FLI1 regulated genes. We reevaluated physical interactions between EWS-FLI1, EWSR1, and RNA Pol II, and used a cross-linking-based strategy to investigate protein assemblies associated with the proteins. The LC domain of EWS-FLI1 was required for the assemblies observed to form in cells. These results offer new insights into a protein assembly that may enable EWS-FLI1 to bind its wide network of protein partners and contribute to regulation of gene expression in Ewing sarcoma.
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影响因子：
48
作者：
Huber W;Carey VJ;Gentleman R;Anders S;Carlson M;Carvalho BS;Bravo HC;Davis S;Gatto L;Girke T;Gottardo R;Hahne F;Hansen KD;Irizarry RA;Lawrence M;Love MI;MacDonald J;Obenchain V;Oleś AK;Pagès H;Reyes A;Shannon P;Smyth GK;Tenenbaum D;Waldron L;Morgan M
通讯作者：
Morgan M
DOI：
10.1074/jbc.m117.800466
发表时间：
2017-11-17
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Lin Y;Currie SL;Rosen MK
通讯作者：
Rosen MK
影响因子：
11.2
作者：
Embree LJ;Azuma M;Hickstein DD
通讯作者：
Hickstein DD
影响因子：
64.8
作者：
Gorthi A;Romero JC;Loranc E;Cao L;Lawrence LA;Goodale E;Iniguez AB;Bernard X;Masamsetti VP;Roston S;Lawlor ER;Toretsky JA;Stegmaier K;Lessnick SL;Chen Y;Bishop AJR
通讯作者：
Bishop AJR
影响因子：
64.8
作者：
Guo, Yang Eric;Manteiga, John C.;Young, Richard A.
通讯作者：
Young, Richard A.