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解析全酵母基因组中转录相关信使核糖核蛋白质量控制组件的动态格局

Deciphering the Dynamic Landscape of Transcription-Associated mRNP Quality Control Components Over the Whole Yeast Genome.

作者信息

Moreau Kévin, Le Dantec Aurélia, Rahmouni A Rachid

机构信息

Centre de Biophysique Moléculaire, UPR 4301 du CNRS, Orléans, France.

出版信息

Methods Mol Biol. 2021;2209:251-265. doi: 10.1007/978-1-0716-0935-4_16.

Abstract

In eukaryotic cells, aberrant mRNPs with processing and packaging defects are targeted co-transcriptionally by a surveillance system that triggers their nuclear retention and ultimately the degradation of their mRNA component by the 3'-5' activity of the exosome-associated exonuclease Rrp6. This mRNP quality control process is stimulated by the NNS complex (Nrd1-Nab3-Sen1), which otherwise mediates termination, processing, and decay of ncRNAs. The process involves also the exosome co-activator TRAMP complex (Trf4-Air2-Mtr4). Here, we describe a genome-wide approach to visualize the dynamic movement and coordination of these quality control components over the yeast chromosomes upon perturbation of mRNP biogenesis. The method provides valuable information on how the surveillance system is precisely coordinated both physically and functionally with the transcription machinery to detect the faulty events during perturbation of mRNP biogenesis. The overview shows also that the gathering of the quality control components over affected mRNA genes takes place at the expense of their commitment to be recruited at ncRNA genomic features, provoking termination and processing defects of ncRNAs.

摘要

在真核细胞中,具有加工和包装缺陷的异常mRNA前体颗粒在转录过程中会被一个监测系统识别,该系统会触发它们在细胞核内的滞留,并最终通过与外切体相关的核酸外切酶Rrp6的3'-5'活性降解其mRNA成分。这种mRNA前体颗粒质量控制过程受到NNS复合物(Nrd1-Nab3-Sen1)的刺激,否则该复合物会介导非编码RNA的终止、加工和降解。这个过程还涉及外切体共激活因子TRAMP复合物(Trf4-Air2-Mtr4)。在这里,我们描述了一种全基因组方法,用于在mRNA前体颗粒生物合成受到干扰时,可视化这些质量控制组件在酵母染色体上的动态移动和协调。该方法提供了关于监测系统如何在物理和功能上与转录机制精确协调,以在mRNA前体颗粒生物合成受到干扰时检测错误事件的有价值信息。概述还表明,质量控制组件在受影响的mRNA基因上的聚集是以牺牲它们在非编码RNA基因组特征处被招募的机会为代价的,从而引发非编码RNA的终止和加工缺陷。

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