Ess2 bridges transcriptional regulators and spliceosomal complexes via distinct interacting domains
Ess2 bridges transcriptional regulators and spliceosomal complexes via distinct interacting domains
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Ess2 通过不同的相互作用域桥接转录调节因子和剪接体复合物
DOI：
10.1016/j.bbrc.2018.02.110
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发表时间：
2018
期刊：
影响因子：
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通讯作者：
Makoto Makishima
中科院分区：
文献类型：
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作者：
Ichiro Takada;Megumi Tsuchiya;Kaori Yanaka;Shinya;Hidano;Sayuri Takahashi;Takashi Kobayashi;Hidesato Ogawa;Sinichi Nakagawa;Makoto Makishima
转录和前体mRNA剪接是涉及转录辅助调节因子的复杂的耦合过程。Dgcr 14是一种核蛋白，可增强视黄酸受体相关孤儿受体γ/γ-t（Rorγ/γt）的转录活性。U2也是剪接体C复合物（含有U2、U 5和U6 snRNA）的一个组成部分。然而，在剪接和转录中起作用的结构域还没有被确定，为了阐明在剪接和转录中的作用，我们进行了RNA免疫沉淀（RIP）试验来检测与转录因子2相互作用的snRNAs。我们发现，U1和U4 snRNA与U6 snRNA相关。使用BMP 2缺失突变体的实验表明，BMP 2的C-末端缺失突变体（1-399 a. a.）失去了与snRNA结合的能力，而NAT 2的N-末端结构域（1-200 a. a.）有趣的是，使用抗ROR共同抗体的实验表明，Rors也与U4和U6 snRNA相关。在T细胞杂交瘤（68-41细胞）中，Rors 2的敲低消除了剪接体与Rors之间的相互作用。在lncRNA（Rmrp）和Rors之间也发现了α 2依赖性关联。因此，我们建议，THE 2协会与转录因子和剪接体复合物和调制剪接反应与转录因子耦合。
Transcription and pre-mRNA splicing are complex, coupled processes that involve transcriptional co-regulators. Ess2 (also termed Dgcr14) is a nuclear protein that enhances the transcriptional activity of retinoic acid receptor-related orphan receptor gamma/gamma-t (Rorγ/γt). Ess2 is also a component of the spliceosomal C complex (containing U2, U5 and U6 snRNAs). However, the domains in Ess2 that function in splicing and transcription have not been identified.To elucidate the roles of Ess2 in splicing and transcription, we performed RNA immunoprecipitation (RIP) assays to detect Ess2-interacting snRNAs. We found that Ess2 associated with U6 snRNA as well as U1 and U4 snRNAs. Experiments using Ess2 deletion mutants showed that a C-terminus deletion mutant of Ess2 (1-399 a. a.) lost its ability to associate with snRNAs, whereas the N-terminus domain of Ess2 (1-200 a. a.) associated with Rorγ/γt, but not with snRNAs.Interestingly, experiments using anti-ROR common antibody showed that Rors also associated with U4 and U6 snRNAs.Ess2knockdown in a T cell hybridoma (68-41 cells) abrogated the interaction between spliceosomes and Rors. An Ess2-dependent association was also found between an lncRNA (Rmrp) and Rors. We thus propose that Ess2 associates with both transcriptional factors and spliceosomal complexes and modulates splicing reactions coupled with transcription factors.