Chateigner-Boutin Anne-Laure, Hanson Maureen R
Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853, USA.
Mol Cell Biol. 2002 Dec;22(24):8448-56. doi: 10.1128/MCB.22.24.8448-8456.2002.
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.
被子植物细胞器中的RNA编辑会导致转录本中的胞嘧啶(Cs)转变为尿嘧啶(Us)。在大多数体外或体内分析的编辑位点中,已发现编辑位点的胞嘧啶(C)上游约30个核苷酸(nt)和下游10个nt范围内的序列对于正确选择C编辑靶点和编辑效率是必需的,但尚未鉴定出共有序列。在两个转基因烟草品系中,针对所有31个已知的编辑C,测定了携带rpoB - 2或ndhF - 2编辑位点的两种不同小基因的高水平表达对编辑的影响。叶绿体转基因植物中,内源性基因转录本以及其他几个基因转录本中相应的rpoB和ndhF编辑位点的编辑效率均降低。在每个过表达编辑位点紧邻的5'端序列以及受到竞争效应影响的其他基因位点中,可以鉴定出保守核苷酸,尽管rpoB - 2的5'端和ndhF - 2的5'端的保守核苷酸不同。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,这些簇在编辑靶点C的5'端30 nt范围内携带保守核苷酸。这些数据与一个模型一致,即相同的反式作用因子根据编辑位点C紧邻的5'端特定序列识别多个叶绿体或线粒体编辑位点。