Identification of novel guide RNAs from the mitochondria of Trypanosome brucei
Identification of novel guide RNAs from the mitochondria of Trypanosome brucei
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DOI：
10.4161/rna.5.2.6043
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发表时间：
2008-04-01
期刊：
影响因子：
4.1
通讯作者：
Goeringer, H. Ulrich
中科院分区：
文献类型：
--
作者：
Madej, Monika J.;Niemann, Moritz;Goeringer, H. Ulrich
非洲锥虫的大多数线粒体mrna都受到RNA编辑反应的影响，其特征是仅插入和/或删除U核苷酸。该反应产生功能性mrna，并由一种高分子质量酶复合物——编辑体催化。编辑体与一类独特的小非编码，3'-寡尿苷化（oU） rna，即所谓的引导rna （gRNAs）相互作用。向导rna在U缺失/插入反应中起交易模板的作用，因此是反应周期中的关键组分。此外，通过利用不同的grna，可以发生不同的编辑事件，从而扩大寄生虫线粒体中的蛋白质多样性。在这项研究中，我们分析了来自布鲁氏锥虫的小的、非编码的线粒体转录本。通过从大小选择的RNA群体中生成cDNA文库，我们鉴定出51个新的ou -RNA。对于其中29种rna，我们能够预测同源mRNA靶标。通过Northern blot分析，我们验证了其中22个ou - rna的表达，并证明它们具有所有已知的gRNA特征。在这51个假定的grna中，有5个的特征是与其同源的完全编辑的mRNA序列不匹配，这表明它们可以作为替代编辑事件的grna。
The majority of mitochondrial mRNAs in African trypanosomes are subject to an RNA editing reaction, which is characterized by the insertion and/or deletion of U nucleotides only. The reaction creates functional mRNAs and is catalyzed by a high molecular mass enzyme complex, the editosome. Editosomes interact with a unique class of small non-coding, 3'-oligouridylated (oU) RNAs, so-called guide RNAs (gRNAs). Guide RNAs function as transacting templates in the U deletion/insertion reaction and thus, represent key components in the reaction cycle. Furthermore, by utilizing different gRNAs, alternative editing events can take place, thereby expanding the protein diversity in the mitochondria of the parasites. In this study, we have analyzed small, non-coding mitochondrial transcripts from Trypanosoma brucei. By generating cDNA libraries from size-selected RNA populations we identified 51 novel oU-RNAs. For 29 of these RNAs we were able to predict cognate mRNA targets. By Northern blot analysis, we verified the expression of 22 of these oU-RNAs and demonstrate that they share all known gRNA characteristics. Five of these 51 putative gRNAs are characterized by single mismatches to their cognate, fully edited mRNA sequences suggesting that they could act as gRNAs for alternative editing events.