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The Arabidopsis Protein CGL20 Is Required for Plastid 50S Ribosome Biogenesis.
Plant Physiology ( IF 6.5 ) Pub Date : 2020-01-14 , DOI: 10.1104/pp.19.01502
Bennet Reiter 1 , Evgenia Vamvaka 1 , Giada Marino 1 , Tatjana Kleine 1 , Peter Jahns 2 , Cordelia Bolle 1 , Dario Leister 1 , Thilo Rühle 3
Affiliation  

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 (Arabidopsis thaliana; 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 (AtCGL20A and AtCGL20B). Inactivation of these genes in the atcgl20ab 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 atcgl20ab 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.

中文翻译:

质体50S核糖体生物发生需要拟南芥蛋白CGL20。

加工和修饰核糖体RNA(rRNA)或参与核糖体组装的辅助因子可促进质体核糖体的生物发生。与它们的细菌和线粒体对应物相比,质体核糖体的生物发生尚不甚为人所知,到目前为止,几乎没有描述辅助因子。在这项研究中,我们报告了拟南芥(Arabidopsis thaliana; AtCGL20)中仅绿色系20(CGL20)中保守的功能表征,这是一种富含Pro的10kD蛋白,靶向线粒体和叶绿体。在拟南芥中,CGL20由具有高度序列相似性的节段重复基因(AtCGL20A和AtCGL20B)编码。atcgl20ab突变体中这些基因的失活导致可见的绿色表型和低温下的生长停滞。叶绿体蛋白质组,在atcgl20ab突变体中,色素组成和光合性能受到显着影响。AtCGL20的缺失会损害质体翻译,扰乱23S rRNA中隐性断裂的形成,并导致叶绿体基质提取物的高分子质量分数中50S核糖体亚基异常积累。此外,即使在RNase处理基质提取物后,AtCGL20A-eGFP融合蛋白也与Suc密度梯度中的50S核糖体亚基竞争。因此,我们建议AtCGL20参与质体中50S核糖体亚基生物发生的后期阶段,该作用可能是由于质体核糖体结构差异而在绿色谱系中演变的。AtCGL20的缺失会损害质体翻译,扰乱23S rRNA中隐性断裂的形成,并导致叶绿体基质提取物的高分子质量分数中50S核糖体亚基异常积累。此外,即使在RNase处理基质提取物后,AtCGL20A-eGFP融合蛋白也与Suc密度梯度中的50S核糖体亚基竞争。因此,我们建议AtCGL20参与质体中50S核糖体亚基生物发生的后期阶段,该作用可能是由于质体核糖体结构差异而在绿色谱系中演变的。AtCGL20的缺失会损害质体翻译,扰乱23S rRNA中隐性断裂的形成,并导致叶绿体基质提取物的高分子质量分数中50S核糖体亚基异常积累。此外,即使在RNase处理基质提取物后,AtCGL20A-eGFP融合蛋白也与Suc密度梯度中的50S核糖体亚基竞争。因此,我们建议AtCGL20参与质体中50S核糖体亚基生物发生的后期阶段,该作用可能是由于质体核糖体结构差异而在绿色谱系中演变的。即使在RNase处理基质提取物后,AtCGL20A-eGFP融合蛋白仍以Suc密度梯度与50S核糖体亚基竞争。因此,我们建议AtCGL20参与质体中50S核糖体亚基生物发生的后期阶段,该作用可能是由于质体核糖体结构差异而在绿色谱系中演变的。即使在RNase处理基质提取物后,AtCGL20A-eGFP融合蛋白仍以Suc密度梯度与50S核糖体亚基竞争。因此,我们建议AtCGL20参与质体中50S核糖体亚基生物发生的后期阶段,该作用可能是由于质体核糖体结构差异而在绿色谱系中演变的。
更新日期:2020-03-03
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