Center for Gene Research, Nagoya University, Nagoya, Japan.
Graduate School of Informatics, Nagoya University, Nagoya, Japan.
Genes Cells. 2022 Apr;27(4):293-304. doi: 10.1111/gtc.12928. Epub 2022 Mar 9.
Pentatricopeptide repeat (PPR) proteins are involved in RNA metabolism and also play a role in posttranscriptional regulation during plant organellar gene expression. Although a hundred of PPR proteins exist in the moss Physcomitrium patens, their functions are not fully understood. Here, we report the function of P-class PPR protein PpPPR_32 in P. patens. A transient expression assay using green fluorescent protein demonstrated that the N-terminal region of PpPPR_32 functions as a chloroplast-targeting transit peptide, indicating that PpPPR_32 is localized in chloroplasts. PpPPR_32 knockout mutants grew autotrophically but with reduced protonema growth and the poor formation of photosystem I (PSI) complexes. Quantitative real-time reverse transcription-polymerase chain reaction and RNA gel blot hybridization analyses revealed a significant reduction in the transcript level of the psaC gene encoding the iron sulfur protein of PSI but no alteration to the transcript levels of other PSI genes. This suggests that PpPPR_32 is specifically involved in the expression level of the psaC gene. Our results indicate that PpPPR_32 is essential for the accumulation of psaC transcript and PSI complexes.
五肽重复(PPR)蛋白参与 RNA 代谢,在植物细胞器基因表达的转录后调控中也发挥作用。虽然在藓类植物Physcomitrium patens 中有一百种 PPR 蛋白存在,但它们的功能尚未完全了解。在这里,我们报告了 P 类 PPR 蛋白 PpPPR_32 在 P. patens 中的功能。使用绿色荧光蛋白的瞬时表达分析表明,PpPPR_32 的 N 端区域作为叶绿体靶向转运肽起作用,表明 PpPPR_32 定位于叶绿体中。PpPPR_32 敲除突变体能够自养生长,但原丝体生长减少,且 PSI 复合物的形成不良。定量实时逆转录聚合酶链反应和 RNA 凝胶印迹杂交分析显示,编码 PSI 铁硫蛋白的 psaC 基因的转录本水平显著降低,但其他 PSI 基因的转录本水平没有改变。这表明 PpPPR_32 特异性参与 psaC 基因的表达水平。我们的结果表明,PpPPR_32 对于 psaC 转录本和 PSI 复合物的积累是必需的。