Wheeler Emily C, Washburn Michael C, Major Francois, Rusch Douglas B, Hundley Heather A
a Medical Sciences Program ; Indiana University ; Bloomington , IN USA.
RNA Biol. 2015;12(2):162-74. doi: 10.1080/15476286.2015.1017220.
ADARs (Adenosine deaminases that act on RNA) "edit" RNA by converting adenosines to inosines within double-stranded regions. The primary targets of ADARs are long duplexes present within noncoding regions of mRNAs, such as introns and 3' untranslated regions (UTRs). Because adenosine and inosine have different base-pairing properties, editing within these regions can alter splicing and recognition by small RNAs. However, despite numerous studies identifying multiple editing sites in these genomic regions, little is known about the extent to which editing sites co-occur on individual transcripts or the functional output of these combinatorial editing events. To begin to address these questions, we performed an ultra-deep sequencing analysis of 4 Caenorhabditis elegans 3' UTRs that are known ADAR targets. Synchronous editing events were determined for the long duplexes in vivo. Furthermore, the validity of each editing event was confirmed by sequencing the same regions of mRNA from worms that lack A-to-I editing. This analysis identified a large number of editing sites that can occur within each 3' UTR, but interestingly, each individual transcript contained only a small fraction of these A-to-I editing events. In addition, editing patterns were not random, indicating that an editing event can affect the efficiency of editing at subsequent adenosines. Furthermore, we identified specific sites that can be both positively and negatively correlated with additional sites leading to mutually exclusive editing patterns. These results suggest that editing in noncoding regions is selective and hyper-editing of cellular RNAs is rare.
作用于RNA的腺苷脱氨酶(ADARs)通过在双链区域将腺苷转化为肌苷来“编辑”RNA。ADARs的主要作用靶点是mRNA非编码区域内的长双链,如内含子和3'非翻译区(UTR)。由于腺苷和肌苷具有不同的碱基配对特性,这些区域内的编辑可以改变剪接以及小RNA的识别。然而,尽管有大量研究在这些基因组区域鉴定出多个编辑位点,但对于单个转录本上编辑位点共现的程度以及这些组合编辑事件的功能输出却知之甚少。为了开始解决这些问题,我们对4个已知为ADAR作用靶点的秀丽隐杆线虫3'UTR进行了超深度测序分析。在体内确定了长双链的同步编辑事件。此外,通过对缺乏A到I编辑的线虫的mRNA相同区域进行测序,证实了每个编辑事件的有效性。该分析在每个3'UTR内鉴定出大量可能出现的编辑位点,但有趣的是,每个单独的转录本只包含这些A到I编辑事件的一小部分。此外,编辑模式并非随机,这表明一个编辑事件可以影响后续腺苷的编辑效率。此外,我们鉴定出了与其他位点呈正相关和负相关的特定位点,导致相互排斥的编辑模式。这些结果表明非编码区域的编辑具有选择性,细胞RNA的过度编辑很少见。