Laboratoire de Microbiologie Cellulaire et Moléculaire et Pathogénicité (MCMP), UMR5234 CNRS- Université Victor Segalen Bordeaux2. 146 rue Léo Saignat 33076 Bordeaux Cedex, France.
Nucleic Acids Res. 2010 Nov;38(20):7112-21. doi: 10.1093/nar/gkq591. Epub 2010 Jul 8.
Most plant mitochondria messenger RNAs (mRNAs) undergo editing through C-to-U conversions located mainly in exon sequences. However, some RNA editing events are found in non-coding regions at critical positions in the predicted secondary and tertiary structures of introns, suggesting that RNA editing could be important for splicing. Here, we studied the relationships between editing and splicing of the mRNA encoding the ribosomal protein S10 (rps10), which has a group II intron and five editing sites. Two of them, C2 and C3, predicted to stabilize the folded structure of the intron necessary for splicing, were studied by using rps10 mutants introduced into isolated potato mitochondria by electroporation. While mutations of C2 involved in EBS2/IBS2 interactions did not affect splicing, probably by the presence of an alternative EBS2' region in domain I of the intron, the edition of site C3 turned out to be critical for rps10 mRNA splicing; only the edited (U) form of the transcript was processed. Interestingly, RNA editing was strongly reduced in transcripts from two different intronless genes, rps10 from potato and cox2 from wheat, suggesting that efficient RNA processing may require a close interaction of factors engaged in different maturation processes. This is the first report linking editing and splicing in conditions close to the in vivo situation.
大多数植物线粒体信使 RNA(mRNA)通过主要位于外显子序列中的 C 到 U 转换进行编辑。然而,一些 RNA 编辑事件发生在预测的内含子二级和三级结构中关键位置的非编码区域,这表明 RNA 编辑对于剪接可能很重要。在这里,我们研究了核糖体蛋白 S10(rps10)mRNA 的编辑和剪接之间的关系,rps10 编码物具有一个 II 组内含子和五个编辑位点。其中两个,C2 和 C3,预测稳定剪接所需的内含子折叠结构,通过电穿孔将 rps10 突变体引入分离的土豆线粒体来研究。虽然 C2 突变不影响剪接,可能是由于内含子 I 域中存在替代的 EBS2' 区域,但 C3 位点的编辑对于 rps10 mRNA 剪接至关重要;只有转录本的编辑(U)形式被加工。有趣的是,来自两个不同无内含子基因(土豆的 rps10 和小麦的 cox2)的转录本中的 RNA 编辑强烈减少,这表明有效的 RNA 加工可能需要参与不同成熟过程的因子之间的紧密相互作用。这是首次在接近体内情况的条件下将编辑与剪接联系起来的报告。