Colas des Francs-Small Catherine, Vincis Pereira Sanglard Lilian, Small Ian
Australian Research Council Centre of Excellence in Plant Energy Biology, School of Molecular Sciences, The University of Western Australia, Crawley, WA, 6009, Australia.
Commun Biol. 2018 Oct 11;1:166. doi: 10.1038/s42003-018-0166-8. eCollection 2018.
Mitochondrial genes encode key components of the cellular energy machinery, but their genetic analysis is difficult or impossible in most organisms (including plants) because of the lack of viable transformation approaches. We report here a method to block the expression of the mitochondrial gene encoding a subunit of respiratory complex I in , via the modification of the specificity of the RNA-binding protein RNA PROCESSING FACTOR 2 (RPF2). We show that the modified RPF2 binds and specifically induces cleavage of RNA, almost eliminating expression of the Nad6 protein and consequently complex I accumulation and activity. To our knowledge, this is the first example of a targeted block in expression of a specific mitochondrial transcript by a custom-designed RNA-binding protein. This opens the path to reverse genetics studies on mitochondrial gene functions and leads to potential applications in agriculture.
线粒体基因编码细胞能量机制的关键组成部分，但由于缺乏可行的转化方法，在大多数生物（包括植物）中对其进行遗传分析都很困难甚至无法进行。我们在此报告一种方法，通过修饰RNA结合蛋白RNA加工因子2（RPF2）的特异性，来阻断拟南芥中编码呼吸复合体I一个亚基的线粒体基因的表达。我们表明，修饰后的RPF2结合并特异性诱导拟南芥RNA的切割，几乎消除了Nad6蛋白的表达，从而导致复合体I的积累和活性降低。据我们所知，这是通过定制设计的RNA结合蛋白靶向阻断特定线粒体转录本表达的首个实例。这为线粒体基因功能的反向遗传学研究开辟了道路，并在农业领域具有潜在应用价值。
