Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), 78000, Versailles, France.
Plant Physiol. 2022 Aug 29;190(1):669-681. doi: 10.1093/plphys/kiac309.
Gene expression in plant mitochondria is predominantly governed at the post-transcriptional level and relies mostly on nuclear-encoded proteins. However, the protein factors involved and the underlying molecular mechanisms are still not well understood. Here, we report on the function of the MITOCHONDRIAL STABILITY FACTOR 3 (MTSF3) protein, previously named EMBRYO DEFECTIVE 2794 (EMB2794), and show that it is essential for accumulation of the mitochondrial NADH dehydrogenase subunit 2 (nad2) transcript in Arabidopsis (Arabidopsis thaliana) but not for splicing of nad2 intron 2 as previously proposed. The MTSF3 gene encodes a pentatricopeptide repeat protein that localizes in the mitochondrion. An MTSF3 null mutation induces embryonic lethality, but viable mtsf3 mutant plants can be generated through partial complementation with the developmentally regulated ABSCISIC ACID INSENSITIVE3 promoter. Genetic analyses revealed growth retardation in rescued mtsf3 plants owing to the specific destabilization of mature nad2 mRNA and a nad2 precursor transcript bearing exons 3 to 5. Biochemical data demonstrate that MTSF3 protein specifically binds to the 3' terminus of nad2. Destabilization of nad2 mRNA induces a substantial decrease in complex I assembly and activity and overexpression of the alternative respiratory pathway. Our results support a role for MTSF3 protein in protecting two nad2 transcripts from degradation by mitochondrial exoribonucleases by binding to their 3' extremities.
植物线粒体中的基因表达主要在转录后水平上进行,并主要依赖于核编码蛋白。然而,涉及的蛋白质因子和潜在的分子机制仍未得到很好的理解。在这里,我们报告了先前命名为胚胎缺陷 2794(EMB2794)的线粒体稳定因子 3(MTSF3)蛋白的功能,并表明它对于拟南芥中线粒体 NADH 脱氢酶亚基 2(nad2)转录本的积累是必不可少的,但对于之前提出的 nad2 内含子 2 的剪接不是必需的。MTSF3 基因编码一种五肽重复蛋白,定位于线粒体中。MTSF3 基因的缺失突变会导致胚胎致死,但通过与发育调节的脱落酸不敏感 3 启动子的部分互补,可以产生存活的 mtsf3 突变体植物。遗传分析显示, rescued mtsf3 植物的生长受到抑制,这是由于成熟 nad2 mRNA 和带有外显子 3 至 5 的 nad2 前体转录本的特异性不稳定所致。生化数据表明,MTSF3 蛋白特异性地结合 nad2 的 3' 末端。nad2 mRNA 的不稳定诱导复合物 I 组装和活性的显著下降,并过表达替代呼吸途径。我们的结果支持 MTSF3 蛋白通过结合其 3' 末端来保护两个 nad2 转录本免受线粒体核糖核酸外切酶降解的作用。