PEPseq量化了转录组范围内的蛋白占用变化,并揭示了转录应激后的选择性翻译抑制
Jakob Trendel1,2, Etienne Boileau3,4, Marco Jochem2
1European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
Nucleic acids research
|July 3, 2023
概括
我们开发了PEPseq,这是一种新的RNA测序方法,用于绘制跨转录组的蛋白质相互作用图. 这揭示了细胞应激期间蛋白质与信使RNA结合的变化,影响了基因调节.
科学领域:
- 分子生物学分子生物学
- 遗传学 遗传学是一种遗传学.
- 生物化学 生物化学
背景情况:
- 转录后基因调节对于细胞反应至关重要.
- 了解RNA结合蛋白的动态是解读基因调节的关键.
- 现有的方法缺乏对蛋白质占用的转录组范围的覆盖.
研究的目的:
- 建立一种用于对RNA的蛋白占用量进行全转录组监测的新方法.
- 为了研究在矿诱导的转化应激过程中蛋白质-RNA相互作用的变化.
- 为了确定受改变蛋白质结合在压力下影响的特定RNA区域和mRNA.
主要方法:
- 开发了用于RNA测序的增强下拉器 (PEPseq).
- 使用代谢性RNA标记4-thiouridine (4SU) 进行光诱导的交叉链接.
- 采用N-hydroxysuccinimide (NHS) 化学物质来分离与蛋白质交联的RNA片段.
主要成果:
- PEPseq成功地监测了整个转录组的蛋白质占用.
- 在矿压力期间观察到特定mRNA的编码区域中蛋白质相互作用的增加.
- 在那些相互作用增加的人中,鉴定了编码细胞质核糖体蛋白的mRNA.
- 定量蛋白质组学证实了这些mRNA在压力恢复过程中被抑制的翻译.
结论:
- PEPseq是一个强大的发现平台,用于对转录后调节的公正调查.
- 化物诱导的压力会改变特定mRNA的蛋白占用率,包括对核糖体蛋白质的mRNA.
- 这些变化与压抑的翻译相关,突出了压力下的调节机制.
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