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在马铃薯线粒体中表达的落叶松未编辑tRNA前体的命运

Fate of a larch unedited tRNA precursor expressed in potato mitochondria.

作者信息

Placido Antonio, Gagliardi Dominique, Gallerani Raffaele, Grienenberger Jean-Michel, Maréchal-Drouard Laurence

机构信息

Dipartimento di Biochimica e Biologia Molecolare, Universita' di Bari, CNR, Via Orabona 4, 70126 Bari, Italy.

出版信息

J Biol Chem. 2005 Sep 30;280(39):33573-9. doi: 10.1074/jbc.M505269200. Epub 2005 Aug 1.

Abstract

In higher plant mitochondria, post-transcriptional C to U conversion known as editing mostly affects mRNAs. However, three tRNAs were also shown to be edited. Among them, three editing sites were identified in larch mitochondrial tRNA(His). We have previously shown that only the edited version can undergo maturation in vitro. In this paper, we introduced via direct DNA uptake the edited or unedited version of larch mitochondrial trnH into isolated potato mitochondria and expressed them under the control of potato mitochondrial 18 S rRNA promoter. As expected, the edited form of larch mitochondrial tRNA(His) precursor was processed in the isolated organelles. By contrast, no mature tRNA(His) was detected when using the unedited version of trnH. However, precursor molecules could be characterized by reverse transcription-PCR. These data demonstrate that the potato mitochondrial editing machinery is not able to recognize these "foreign" editing sites and confirm that these unedited tRNA precursor molecules are not correctly processed in organello. As a consequence, the fate of these RNA precursor molecules is likely to be degradation. Indeed, we detected by PCR two 3'-end truncated precursor RNAs. Interestingly, both RNA species exhibit poly(A) tails, a hallmark of degradation in plant mitochondria. Taken together, these data suggest that, in plant mitochondria, a defective unedited RNA precursor that cannot be processed to give a mature stable tRNA, is degraded through a polyadenylation-dependent pathway.

摘要

在高等植物线粒体中,被称为编辑的转录后C到U的转换主要影响mRNA。然而,也有三种tRNA被证明会发生编辑。其中,在落叶松线粒体tRNA(His)中鉴定出三个编辑位点。我们之前已经表明,只有经过编辑的版本才能在体外进行成熟。在本文中,我们通过直接摄取DNA将落叶松线粒体trnH的编辑或未编辑版本导入分离的马铃薯线粒体中,并在马铃薯线粒体18 S rRNA启动子的控制下进行表达。正如预期的那样,落叶松线粒体tRNA(His)前体的编辑形式在分离的细胞器中被加工。相比之下,当使用trnH的未编辑版本时,未检测到成熟的tRNA(His)。然而,前体分子可以通过逆转录PCR进行表征。这些数据表明,马铃薯线粒体编辑机制无法识别这些“外来”的编辑位点,并证实这些未编辑的tRNA前体分子在细胞器中没有被正确加工。因此,这些RNA前体分子的命运可能是降解。事实上,我们通过PCR检测到了两种3'端截短的前体RNA。有趣的是,这两种RNA都具有poly(A)尾巴,这是植物线粒体降解的一个标志。综上所述,这些数据表明,在植物线粒体中,有缺陷的未编辑RNA前体无法被加工成成熟稳定的tRNA,而是通过多聚腺苷酸化依赖的途径被降解。

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