Laboratory of Environmental Plant Biotechnology, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumidori-Amamiyamachi, Sendai 981-8555, Japan.
Plant J. 2012 Nov;72(3):450-60. doi: 10.1111/j.1365-313X.2012.05091.x. Epub 2012 Aug 14.
Pentatricopeptide repeat (PPR) proteins are involved in the modification of organelle transcripts. In this study, we investigated the molecular function in rice of the mitochondrial PPR-encoding gene MITOCHONDRIAL PPR25 (MPR25), which belongs to the E subgroup of the PPR family. A Tos17 knockout mutant of MPR25 exhibited growth retardation and pale-green leaves with reduced chlorophyll content during the early stages of plant development. The photosynthetic rate in the mpr25 mutant was significantly decreased, especially under strong light conditions, although the respiration rate did not differ from that of wild-type plants. MPR25 was preferentially expressed in leaves. FLAG-tagged MPR25 accumulated in mitochondria but not in chloroplasts. Direct sequencing revealed that the mpr25 mutant fails to edit a C-U RNA editing site at nucleotide 1580 of nad5, which encodes a subunit of complex I (NADH dehydrogenase) of the respiratory chain in mitochondria. RNA editing of this site is responsible for a change in amino acid from serine to leucine. Recombinant MPR25 directly interacted with the proximal region of the editing site of nad5 transcripts. However, the NADH dehydrogenase activity of complex I was not affected in the mutant. By contrast, genes encoding alternative NADH dehydrogenases and alternative oxidase were up-regulated. The mpr25 mutant may therefore provide new information on the coordinated interaction between mitochondria and chloroplasts.
五肽重复(PPR)蛋白参与细胞器转录物的修饰。在这项研究中,我们研究了属于 PPR 家族 E 亚组的线粒体 PPR 编码基因 MITOCHONDRIAL PPR25(MPR25)在水稻中的分子功能。MPR25 的 Tos17 敲除突变体在植物发育的早期表现出生长缓慢和浅绿色叶片,叶绿素含量降低。尽管呼吸速率与野生型植物没有差异,但 mpr25 突变体的光合速率显著降低,尤其是在强光条件下。MPR25 在叶片中优先表达。标记有 FLAG 的 MPR25 积累在线粒体中,但不在叶绿体中。直接测序表明,mpr25 突变体未能编辑 nad5 第 1580 位核苷酸的 C-U RNA 编辑位点,该位点编码呼吸链复合体 I(NADH 脱氢酶)的亚基。该位点的 RNA 编辑负责将氨基酸从丝氨酸变为亮氨酸。重组 MPR25 直接与 nad5 转录物编辑位点的近端区域相互作用。然而,突变体中复合体 I 的 NADH 脱氢酶活性并未受到影响。相比之下,编码替代 NADH 脱氢酶和替代氧化酶的基因上调。因此,mpr25 突变体可能为线粒体和叶绿体之间的协调相互作用提供新的信息。