在R环调节中与长非编码RNA TUG1 相互作用的蛋白质的全面识别
Jingqi Xie1, Miho M Suzuki1, Kenta Iijima2
1Division of Cancer Biology, Nagoya University Graduate School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya, Aichi 466-8550, Japan.
Journal of biochemistry
|July 14, 2025
概括
氨酸高调基因1 (TUG1),一个长的非编码RNA,与蛋白质相互作用以保持基因组完整性. 这项研究确定了TUG1的身份.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 在RNA生物学,RNA生物学.
背景情况:
- 长非编码RNAs (lncRNAs) 是通过RNA结合蛋白 (RBP) 相互作用的细胞过程的关键调节者.
- 氨酸上调基因1 (TUG1) 是癌症中过度表达的lncRNA,参与R循环解析和基因组完整性维护.
- 图格1的完整的蛋白相互作用场景和应激反应在很大程度上仍然没有特征.
研究的目的:
- 通过使用CRISPR辅助的RNA-蛋白相互作用检测 (CARPID) 来全面识别TUG1相互作用蛋白.
- 为了研究在R循环诱导压力条件下TUG1-蛋白相互作用的变化.
- 通过其蛋白质相互作用,阐明TUG1在基因组维护中的作用.
主要方法:
- 通过CRISPR辅助的RNA-蛋白相互作用检测 (CARPID) 与质谱学相结合.
- 利用了三个不同的单导向RNA (sgRNAs),在HEK293T细胞中准TUG1.
- 在基底和坎プト素 (CPT) 诱导的R循环应激条件下分析了蛋白质相互作用.
主要成果:
- 在基底条件下确定了17种TUG1相互作用蛋白.
- 在CPT治疗时增加到25个相互作用蛋白,表明动态的RBP结合.
- 观察到在压力下识别的蛋白质中存在较大的重叠,这表明保留了R循环反应网络.
- 已确认与已知的R环因子 (如RNA基酶,PARP1和HNRNP) 的相互作用.
结论:
- TUG1与一组定义的蛋白质相互作用,以保持基因组完整性.
- 在压力下,TUG1积极启动R循环调节机制.
- 提供了关于 lncRNA 介导的 R 循环调节及其对基因组稳定性的贡献的新见解.
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