Wang Chuande, Aubé Fabien, Planchard Noelya, Quadrado Martine, Dargel-Graffin Céline, Nogué Fabien, Mireau Hakim
Institut Jean-Pierre Bourgin, INRA, AgroParisTech, CNRS, Université Paris-Saclay, RD10, 78026 Versailles Cedex, France.
Paris-Sud University, Université Paris-Saclay, 91405 Orsay Cedex, France.
Nucleic Acids Res. 2017 Jun 2;45(10):6119-6134. doi: 10.1093/nar/gkx162.
RNA expression in plant mitochondria implies a large number of post-transcriptional events in which transcript processing and stabilization are essential. In this study, we analyzed the function of the Arabidopsis mitochondrial stability factor 2 gene (MTSF2) and show that the encoded pentatricopeptide repeat protein is essential for the accumulation of stable nad1 mRNA. The production of mature nad1 requires the assembly of three independent RNA precursors via two trans-splicing reactions. Genetic analyses revealed that the lack of nad1 in mtsf2 mutants results from the specific destabilization of the nad1 exons 2-3 precursor transcript. We further demonstrated that MTSF2 binds to its 3΄ extremity with high affinity, suggesting a protective action by blocking exoribonuclease progression. By defining the 3΄ end of nad1 exons 2-3 precursor, MTSF2 concomitantly determines the 3΄ extremity of the first half of the trans-intron found at the end of the transcript. Therefore, binding of the MTSF2 protein to nad1 exons 2-3 precursor evolved both to stabilize the transcript and to define a 3΄ extremity compatible with the trans-splicing reaction needed to reconstitute mature nad1. We thus reveal that the range of transcripts stabilized by association with protective protein on their 3΄ end concerns also mitochondrial precursor transcripts.
植物线粒体中的RNA表达意味着大量的转录后事件,其中转录本加工和稳定化至关重要。在本研究中,我们分析了拟南芥线粒体稳定因子2基因(MTSF2)的功能,并表明编码的五肽重复序列蛋白对于稳定的nad1 mRNA积累至关重要。成熟nad1的产生需要通过两个反式剪接反应组装三个独立的RNA前体。遗传分析表明,mtsf2突变体中nad1的缺失是由于nad1外显子2 - 3前体转录本的特异性不稳定所致。我们进一步证明,MTSF2以高亲和力结合其3΄末端,表明通过阻止外切核糖核酸酶的进程起到保护作用。通过确定nad1外显子2 - 3前体的3΄末端,MTSF2同时确定了转录本末端发现的反式内含子前半部分的3΄末端。因此,MTSF2蛋白与nad1外显子2 - 3前体的结合既进化为稳定转录本,又确定了与重组成熟nad1所需的反式剪接反应兼容的3΄末端。我们由此揭示,通过在其3΄末端与保护蛋白结合而稳定的转录本范围也涉及线粒体前体转录本。