P-class Pentatricopeptide Repeat Protein Research Articles

PRECOCIOUS1 (POCO1), a mitochondrial pentatricopeptide repeat protein affects flowering time in Arabidopsis thaliana.

Flowering is a vital developmental shift in plants from vegetative to reproductive phase. The timing of this shift is regulated by various linked genetic pathways including environmental cues and internal regulation. Here we report a role for an Arabidopsis gene, AT1G15480, which encodes a P-class pentatricopeptide repeat (PPR) protein, affecting flowering time. We show that AT1G15480 is localized to mitochondria. An AT1G15480 T-DNA insertion line exhibits an early-flowering phenotype, which is quite a rare phenotype among PPR mutants. The early-flowering phenotype was observed under both long and short days compared with wild type plants. Genetic complementation confirmed the observed phenotype. We therefore named the PPR protein PRECOCIOUS1 (POCO1). poco1 plants showed lower respiration, ATP content and higher accumulation of superoxide. Importantly, the quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis showed that the expression of FLOWERING LOCUS C (FLC), which is a key floral repressor, was strongly downregulated in the poco1. Likewise, the expression level of the FLC positive regulator ABSCISIC ACID-INSENSITIVE 5 (ABI5) was reduced in the poco1. Consistent with the qRT-PCR results, poco1 plants showed reduced sensitivity to abscisic acid compared with wild type with respect to primary root growth and days to flowering. Furthermore, the poco1 mutation enhances the sensitivity to drought stress. Further analysis showed that POCO1 affects mitochondrial RNA editing. Taken together, our data demonstrate a remarkable function of POCO1 in flowering time and the abscisic acid signalling pathway.

PPR Protein BFA2 Is Essential for the Accumulation of the atpH/F Transcript in Chloroplasts.

As a fascinating and complicated nanomotor, chloroplast ATP synthase comprises nine subunits encoded by both the nuclear and plastid genomes. Because of its uneven subunit stoichiometry, biogenesis of ATP synthase and expression of plastid-encoded ATP synthase genes requires assistance by nucleus-encoded factors involved in transcriptional, post-transcriptional, and translational steps. In this study, we report a P-class pentatricopeptide repeat (PPR) protein BFA2 (Biogenesis Factor required for ATP synthase 2) that is essential for accumulation of the dicistronic atpH/F transcript in Arabidopsis chloroplasts. A loss-of-function mutation in BFA2 results in a specific reduction of more than 3/4 of chloroplast ATP synthase, which is likely due to the absence of dicistronic atpH/F transcript. BFA2 protein contains 22 putative PPR motifs and exclusively localizes in the chloroplast. Bioinformatics and Electrophoretic Mobility Shift Assays (EMSA) analysis showed that BFA2 binds to the consensus sequence of the atpF-atpA intergenic region in a sequence-specific manner. However, translation initiation of the atpA was not affected in the bfa2 mutant. Thus, we propose that the chloroplast PPR protein BFA2 mainly acts as barrier to prevent the atpH/F transcript degradation by exoribonucleases by binding to the consensus sequence of the atpF-atpA intergenic region.

RNA Processing Factor 7 and Polynucleotide Phosphorylase Are Necessary for Processing and Stability of nad2 mRNA in Arabidopsis Mitochondria

Post-transcriptional maturation of plant mitochondrial transcripts requires several steps. Among these, the generation of mature 5′ ends is still one of the most enigmatic processes. Toward a characterization of proteins involved in 5′ processing of mitochondrial transcripts in Arabidopsis (Arabidopsis thaliana), we now analyzed 5′ maturation of nad2 transcripts. Based on natural genetic variation affecting 5′ ends of nad2 transcripts in ecotype Can-0 and complementation studies we now identified RNA processing factor 7, which takes part in the generation of the 5′ terminus of the mature nad2 mRNA. RPF7 is a relatively short regular P-class pentatricopeptide repeat protein comprising seven canonical P repeats and a single short S repeat. The corresponding allele in Can-0 encodes a truncated version of this protein lacking two C-terminal repeats, which are essential for the function of RPF7. Furthermore we established transgenic plants expressing artifical microRNAs targeting the mitochondrial polynucleotide phosphorylase (PNPase), which results in substantial reduction of the PNPase mRNA levels and strong knockdown of this gene. Detailed quantitative studies of 5′ and 3′ extended nad2 precursor RNAs in these knockdown plants as well as in the rpf7–1 knockout mutant suggest that 5′ processing contributes to the stability of mitochondrial transcripts in plants.