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无义介导的 mRNA 降解控制了渗透压胁迫下酵母核糖体蛋白前体 mRNA 水平的变化。

Nonsense-mediated mRNA decay controls the changes in yeast ribosomal protein pre-mRNAs levels upon osmotic stress.

机构信息

Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Biológicas Universitat de València, Valencia, Spain.

出版信息

PLoS One. 2013 Apr 19;8(4):e61240. doi: 10.1371/journal.pone.0061240. Print 2013.

Abstract

The expression of ribosomal protein (RP) genes requires a substantial part of cellular transcription, processing and translation resources. Thus, the RP expression must be tightly regulated in response to conditions that compromise cell survival. In Saccharomyces cerevisiae cells, regulation of the RP gene expression at the transcriptional, mature mRNA stability and translational levels during the response to osmotic stress has been reported. Reprogramming global protein synthesis upon osmotic shock includes the movement of ribosomes from RP transcripts to stress-induced mRNAs. Using tiling arrays, we show that osmotic stress yields a drop in the levels of RP pre-mRNAs in S. cerevisiae cells. An analysis of the tiling array data, together with transcription rates data, shows a poor correlation, indicating that the drop in the RP pre-mRNA levels is not merely a result of the lowered RP transcription rates. A kinetic study using quantitative RT-PCR confirmed the decrease in the levels of several RP-unspliced transcripts during the first 15 minutes of osmotic stress, which seems independent of MAP kinase Hog1. Moreover, we found that the mutations in the components of the nonsense-mediated mRNA decay (NMD), Upf1, Upf2, Upf3 or in exonuclease Xrn1, eliminate the osmotic stress-induced drop in RP pre-mRNAs. Altogether, our results indicate that the degradation of yeast RP unspliced transcripts by NMD increases during osmotic stress, and suggest that this might be another mechanism to control RP synthesis during the stress response.

摘要

核糖体蛋白 (RP) 基因的表达需要细胞转录、加工和翻译资源的很大一部分。因此,RP 的表达必须根据危及细胞存活的条件进行严格调节。在酿酒酵母细胞中,已有报道称在应对渗透压胁迫时,RP 基因表达在转录、成熟 mRNA 稳定性和翻译水平上受到调节。渗透压休克后全局蛋白质合成的重编程包括核糖体从 RP 转录本向应激诱导的 mRNA 的移动。使用平铺阵列,我们表明渗透压胁迫会导致酿酒酵母细胞中 RP 前体 mRNA 水平下降。对平铺阵列数据的分析以及转录率数据表明相关性较差,表明 RP 前体 mRNA 水平的下降不仅仅是 RP 转录率降低的结果。使用定量 RT-PCR 的动力学研究证实,在渗透压胁迫的最初 15 分钟内,几种 RP 未剪接转录本的水平下降,这似乎独立于 MAP 激酶 Hog1。此外,我们发现,无意义介导的 mRNA 降解 (NMD) 组件、Upf1、Upf2、Upf3 或外切酶 Xrn1 的突变消除了渗透压胁迫诱导的 RP 前体 mRNA 下降。总之,我们的结果表明,NMD 对酵母 RP 未剪接转录本的降解在渗透压胁迫期间增加,并表明这可能是应激反应中控制 RP 合成的另一种机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9700/3631235/5cf8b8aac341/pone.0061240.g001.jpg

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