An in vitro RNA editing system from cauliflower mitochondria: Editing site recognition parameters can vary in different plant species
An in vitro RNA editing system from cauliflower mitochondria: Editing site recognition parameters can vary in different plant species
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DOI：
10.1261/rna.2740905
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发表时间：
2005-10-01
期刊：
影响因子：
4.5
通讯作者：
Brennicke, A
中科院分区：
文献类型：
--
作者：
Neuwirt, J;Takenaka, M;Brennicke, A
开花植物线粒体中400个RNA编辑位点大部分位于mrna中。因此，同源位点的序列邻近在不同物种之间是高度保守的，并且可能被同样保守的转录因子识别。为了研究对序列变异的进化适应，我们分析了豌豆和花椰菜两个物种中上游序列不同的编辑位点的识别元件。这种变异在选择的位点上是可以容忍的，因为上游的顺式元件到达mRNA的5'-UTR。为了比较豌豆和花椰菜线粒体的顺式识别特征，我们开发了一种新的花椰菜体外RNA编辑系统。缺失和突变模板rna的体外编辑实验表明，这两个物种的主要识别元件都位于保守序列内。然而，在花椰菜中，必需的上游核苷酸比在豌豆中延伸得更远。对单核苷酸突变的深入分析揭示了编辑位点的临界间隔和特定的识别元件，并表明+1核苷酸的同源性在花椰菜中很重要，而在豌豆中并不重要。
Most of the 400 RNA editing sites in flowering plant mitochondria are found in mRNAs. Consequently, the sequence vicinities of homologous sites are highly conserved between different species and are presumably recognized by likewise conserved transfactors. To investigate the evolutionary adaptation to sequence variation, we have now analyzed the recognition elements of an editing site with divergent upstream sequences in the two species pea and cauliflower. This variation is tolerated at the site selected, because the upstream cis-elements reach into the 5'-UTR of the mRNA. To compare cis-recognition features in pea and cauliflower mitochondria, we developed a new in vitro RNA editing system for cauliflower. in vitro editing assays with deleted and mutated template RNAs show that the major recognition elements for both species are located within the conserved sequence. In cauliflower, however, the essential upstream nucleotides extend further upstream than they do in pea. In-depth analysis of single-nucleotide mutations reveals critical spacing of the editing site and the specific recognition elements, and shows that the +1 nucleotide identity is important in cauliflower, but not in pea.