Plant Molecular Biology Faculty of Biology I, Ludwig-Maximilians-Universität, D-82152 Planegg-Martinsried, Germany.
Institute of Plant Biochemistry, Heinrich-Heine University, 40225 Duesseldorf, Germany.
Plant Physiol. 2020 Mar;182(3):1222-1238. doi: 10.1104/pp.19.01502. Epub 2020 Jan 14.
Biogenesis of plastid ribosomes is facilitated by auxiliary factors that process and modify ribosomal RNAs (rRNAs) or are involved in ribosome assembly. In comparison with their bacterial and mitochondrial counterparts, the biogenesis of plastid ribosomes is less well understood, and few auxiliary factors have been described so far. In this study, we report the functional characterization of CONSERVED ONLY IN THE GREEN LINEAGE20 (CGL20) in Arabidopsis (; AtCGL20), which is a Pro-rich, ∼10-kD protein that is targeted to mitochondria and chloroplasts. In Arabidopsis, CGL20 is encoded by segmentally duplicated genes of high sequence similarity ( and ). Inactivation of these genes in the mutant led to a visible virescent phenotype and growth arrest at low temperature. The chloroplast proteome, pigment composition, and photosynthetic performance were significantly affected in mutants. Loss of AtCGL20 impaired plastid translation, perturbing the formation of a hidden break in the 23S rRNA and causing abnormal accumulation of 50S ribosomal subunits in the high-molecular-mass fraction of chloroplast stromal extracts. Moreover, AtCGL20A-eGFP fusion proteins comigrated with 50S ribosomal subunits in Suc density gradients, even after RNase treatment of stromal extracts. Therefore, we propose that AtCGL20 participates in the late stages of the biogenesis of 50S ribosomal subunits in plastids, a role that presumably evolved in the green lineage as a consequence of structural divergence of plastid ribosomes.
质体核糖体的生物发生是由辅助因子促进的,这些辅助因子可以处理和修饰核糖体 RNA(rRNA)或参与核糖体组装。与细菌和线粒体的对应物相比,质体核糖体的生物发生还不太了解,迄今为止,描述的辅助因子很少。在这项研究中,我们报告了拟南芥中保守仅在绿色谱系 20(CGL20)(AtCGL20)的功能特征,它是一种富含脯氨酸、约 10kDa 的蛋白质,靶向线粒体和叶绿体。在拟南芥中,CGL20 由高度相似序列的分段重复基因编码(和)。这些基因在 突变体中的失活导致可见的黄绿色表型和低温生长停滞。叶绿体蛋白质组、色素组成和光合作用性能在 突变体中受到显著影响。AtCGL20 的缺失损害了质体翻译,扰乱了 23S rRNA 中的隐藏断裂的形成,并导致异常积累 50S 核糖体亚基在叶绿体基质提取物的高分子质量部分。此外,AtCGL20A-eGFP 融合蛋白与 Suc 密度梯度中的 50S 核糖体亚基共迁移,甚至在对基质提取物进行 RNase 处理后也是如此。因此,我们提出 AtCGL20 参与质体中 50S 核糖体亚基生物发生的晚期阶段,这种作用可能是由于质体核糖体结构的分化而在绿色谱系中进化而来的。