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OsPPR939, a nad5 splicing factor, is essential for plant growth and pollen development in rice
Saturday, 2021/03/13 | 08:03:40

Peng Zheng, Yujun Liu, Xuejiao Liu, Yuqing Huang, Feng Sun, Wenyi Wang, Hao Chen, Mehmood Jan, Cuicui Zhang, Yue Yuan, Bao-Cai Tan, Hao Du & Jumin Tu

Theoretical and Applied Genetics March 2021; vol. 134:  923–940

Key message

P-subfamily PPR protein OsPPR939, which can be phosphorylated by OsS6K1, regulates plant growth and pollen development by involving in the splicing of mitochondrial nad5 introns 1, 2, and 3.

Abstract

In land plants, pentatricopeptide repeat (PPR) proteins play key roles in mitochondrial group II intron splicing, but how these nucleus-encoded proteins are imported into mitochondria is unknown. To date, a few PPR proteins have been characterized in rice (Oryza sativa). Here, we demonstrate that the mitochondrion-localized P-subfamily PPR protein OsPPR939 is required for the splicing of nad5 introns 1, 2, and 3 in rice. Complete knockout or partial disruption of OsPPR939 function resulted in different degrees of growth retardation and pollen sterility. The dramatically reduced splicing efficiency of these introns in osppr939-4 and osppr939-5 led to reduced mitochondrial complex I abundance and activity and enhanced expression of alternative respiratory pathway genes. Complementation with OsPPR939 rescued the defective plant morphology of osppr939-4 and restored its decreased splicing efficiency of nad5 introns 1, 2, and 3. Therefore, OsPPR939 plays crucial roles in plant growth and pollen development by splicing mitochondrial nad5 introns 1, 2, and 3. More importantly, the 12th amino acid Ser in the N-terminal targeting sequence of OsPPR939 is phosphorylated by OsS6K1, and truncated OsPPR939 with a non-phosphorylatable S12A mutation in its presequence could not be imported into mitochondria, suggesting that phosphorylation of this amino acid plays an important role in the mitochondrial import of OsPPR939. To our knowledge, the 12th residue Ser on OsPPR939 is the first experimentally proven phosphorylation site in PPR proteins. Our results provide a basis for investigating the regulatory mechanism of PPR proteins at the post-translational level.

 

See: https://link.springer.com/article/10.1007/s00122-020-03742-6

 

Figure 4: Complementation of osppr939-4 and tang2. a Complementation with OsPPR939pro:OsPPR939 rescued the defective phenotypes of osppr939-4. 1 Shows a wild type (WT) plant, osppr939-4 plant, and OsPPR939pro:OsPPR939-complemented (osppr939-4COM) plant expressing the wild type OsPPR939 gene. 2 Shows I2-KI staining of pollen grains from wild type, osppr939-4, and osppr939-4COM plants. 3 Shows RT-PCR analysis of mature nad5 transcripts from wild type, osppr939-4, and three complemented lines. OsGAPDH was used as a loading control. Bars = 10 cm (1), 100 μm (2). b OsPPR939pro:OsPPR939 rescued the defective phenotypes of tang2. 1 Shows the structure of the Arabidopsis mitochondrial nad5 gene. The positions of the primers used in 2 are shown. 2 Shows RT-PCR analysis of Arabidopsis nad5 intron 3 spliced transcripts in Col-0, tang2, and three complemented lines expressing OsPPR939. 3 Shows a Col-0 plant, tang2 plant, and complemented tang2COM plant. Col-0, Columbia-0. Bars = 1 cm.

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