The moss pentatricopeptide repeat protein with a DYW domain is responsible for RNA editing of mitochondrial ccmFc transcript
The moss pentatricopeptide repeat protein with a DYW domain is responsible for RNA editing of mitochondrial ccmFc transcript
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DOI：
10.1111/j.1365-313x.2010.04175.x
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发表时间：
2010-05-01
期刊：
影响因子：
7.2
通讯作者：
Sugita, Mamoru
中科院分区：
文献类型：
--
作者：
Tasaki, Eiji;Hattori, Mitsuru;Sugita, Mamoru
在大多数陆生植物中，RNA编辑经常发生在许多细胞器转录本中，但对细胞器RNA编辑过程的分子机制知之甚少。在这项研究中，我们的特点是小立碗藓PpPPR_71基因所需的ccmFc转录的RNA编辑。该转录物含有两个RNA编辑位点，ccmF-1和ccmF-2，它们被18个核苷酸分开。ccmFc的互补DNA序列分析表明，在ccmF-1位点的RNA编辑发生在ccmF-2编辑之前。ccmF-2下游位点的RNA编辑通过PpPPR_71基因的破坏而特异性受损，该基因编码具有17个五肽重复基序和C末端DYW结构域的多肽。在大肠杆菌中表达的重组PpPPR_71蛋白与含有ccmF-2编辑位点的46个核苷酸的序列特异性结合。当使用在ccmF-1下编辑的RNA时，重组PpPPR_71的结合亲和力最强。此外，DYW结构域还与周围的ccmF-2编辑位点结合。我们的结论是，PpPPR_71是一个RNA结合蛋白，作为一个位点识别因子在线粒体RNA编辑。
P>In most land plants RNA editing frequently occurs in many organelle transcripts, but little is known about the molecular mechanisms of the organelle RNA editing process. In this study, we have characterized the Physcomitrella patens PpPPR_71 gene that is required for RNA editing of the ccmFc transcript. This transcript harbors two RNA editing sites, ccmF-1 and ccmF-2, that are separated by 18 nucleotides. Complementary DNA sequence analysis of ccmFc suggested that RNA editing at the ccmF-1 site occurred before ccmF-2 editing. RNA editing of the ccmF-2 downstream site was specifically impaired by disruption of the PpPPR_71 gene that encodes a polypeptide with 17 pentatricopeptide repeat motifs and a C-terminal DYW domain. The recombinant PpPPR_71 protein expressed in Escherichia coli specifically bound to the 46-nucleotide sequence containing the ccmF-2 editing site. The binding affinity of the recombinant PpPPR_71 was strongest when using the edited RNA at ccmF-1. In addition, the DYW domain also binds to the surrounding ccmF-2 editing site. We conclude that PpPPR_71 is an RNA-binding protein that acts as a site recognition factor in mitochondrial RNA editing.