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Cross-Competition in Transgenic Chloroplasts Expressing Single Editing Sites Reveals Shared cis Elements

机译:表达单个编辑位点的转基因叶绿体中的交叉竞争揭示了共享的顺式元素。

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摘要

RNA editing in organelles of angiosperm plants results in alteration of Cs to Us in transcripts. In most editing sites analyzed in vitro or in vivo, sequences within approximately 30 nucleotides (nt) 5′ and 10 nt 3′ of the edited C have been found to be required for selection of the correct C editing target and for editing efficiency, but no consensus sequences have been identified. The effect of high-level expression of two different minigenes carrying either the rpoB-2 or the ndhF-2 editing site on editing was determined for all 31 known edited Cs in two transgenic tobacco lines. The editing efficiencies of both the corresponding rpoB and ndhF editing sites in the endogenous genes' transcripts and in several other genes' transcripts were reduced in the chloroplast transgenic plants. Conserved nucleotides could be identified in the sequences immediately 5′ of each overexpressed editing site and in the sites in the additional genes that experienced a competition effect, though the conserved nucleotides differ 5′ of rpoB-2 and 5′ of ndhF-2. Inspection of sequences surrounding chloroplast and mitochondrial editing sites reveals that they can be grouped into clusters which carry conserved nucleotides within 30 nt 5′ of the C target of editing. The data are consistent with a model in which the same trans factor recognizes several chloroplast or mitochondrial editing sites, depending on the particular sequence 5′ of the edited C.
机译:被子植物细胞器中的RNA编辑导致转录本中Cs改变为Us。在体外或体内分析的大多数编辑位点中,已发现选择C的正确编辑目标和提高编辑效率需要约30个核苷酸(nt)5'和10 nt 3'内的序列。尚未确定共有序列。对于两个转基因烟草品系中的所有31个已知编辑的C,确定了携带rpoB-2或ndhF-2编辑位点的两个不同小基因高水平表达对编辑的影响。在叶绿体转基因植物中,相应的rpoB和ndhF编辑位点在内源基因转录本和其他几个基因转录本中的编辑效率均降低。保守的核苷酸可以在每个过表达的编辑位点5'的序列中以及在其他具有竞争作用的基因的位点中立即鉴定出来,尽管保守的核苷酸不同于rpoB-2的5'和ndhF-2的5'。对叶绿体和线粒体编辑位点周围序列的检查表明,它们可以被分组为簇,这些簇在C的编辑目标的30 nt 5'内带有保守的核苷酸。数据与模型一致,在模型中,相同的反式因子可识别多个叶绿体或线粒体编辑位点,具体取决于被编辑C的特定序列5'。

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