Protein Abundance Control by Non-coding Antisense Transcription.
Protein Abundance Control by Non-coding Antisense Transcription.
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DOI：
10.1016/j.celrep.2016.05.043
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发表时间：
2016-06-21
期刊：
影响因子：
8.8
通讯作者：
Knop M
中科院分区：
文献类型：
--
作者：
Huber F;Bunina D;Gupta I;Khmelinskii A;Meurer M;Theer P;Steinmetz LM;Knop M
稳定的未注释转录本 (SUT) 是一类非编码 RNA，其中一些在反义方向上与蛋白质编码基因重叠。虽然案例研究报告了其中几种 RNA 的重要调节作用，但它们对重叠基因的蛋白质丰度调节的一般影响尚不清楚。为了测试这一点，我们通过使用单向转录终止子和 GFP 标签，采用无缝基因操作来抑制 162 个酵母基因的反义 SUT。我们发现，仅存在反义 SUT 不足以影响蛋白质丰度，观察到的反义 SUT 的影响与有义转录起始位点重叠相关，而且这种影响通常很弱，导致蛋白质水平降低。反义调节基因显示 H3K4 二甲基化和三甲基化增加，并且噪音水平略低于预期。我们的结果表明反义 RNA 的功能具有基因和条件特异性成分。测量了 188 个基因的反义转录对蛋白质水平的影响 反义对 25% 的基因大多具有较弱的抑制作用 反义调节与启动子重叠和 H3K4 甲基化相关 反义调节基因的噪音水平低于预期 Huber 等人。通过选择性抑制 188 个 GFP 标记基因的反义转录本并量化由此产生的蛋白质表达水平变化，对酵母中反义 RNA 的功能进行了系统研究。作者利用这些信息来识别区分功能性反义 RNA 和非功能性反义 RNA 的特征。
Stable unannotated transcripts (SUTs), some of which overlap protein-coding genes in antisense direction, are a class of non-coding RNAs. While case studies have reported important regulatory roles for several of such RNAs, their general impact on protein abundance regulation of the overlapping gene is not known. To test this, we employed seamless gene manipulation to repress antisense SUTs of 162 yeast genes by using a unidirectional transcriptional terminator and a GFP tag. We found that the mere presence of antisense SUTs was not sufficient to influence protein abundance, that observed effects of antisense SUTs correlated with sense transcript start site overlap, and that the effects were generally weak and led to reduced protein levels. Antisense regulated genes showed increased H3K4 di- and trimethylation and had slightly lower than expected noise levels. Our results suggest that the functionality of antisense RNAs has gene and condition-specific components. The impact of antisense transcription on protein levels is measured for 188 genes Antisense has mostly weak suppressive effects on ∼25% of the genes Regulation by antisense correlates with promoter overlap and H3K4 methylation Antisense-regulated genes have lower than expected noise levels Huber et al. conduct a systematic study of the function of antisense RNAs in yeast by selectively suppressing the antisense transcripts of 188 GFP-tagged genes and quantifying the resulting changes in protein expression levels. The authors use this information to identify features that distinguish functional from non-functional antisense RNAs.