Site-specific binding of a PPR protein defines and stabilizes 50 and 30 mRNA termini in chloroplasts

Chloroplast mRNA populations are characterized by overlapping transcripts derived by processing from polycistronic precursors. The mechanisms and functional significance of these processing events are poorly understood. We describe a pentatricopeptide repeat (PPR) protein, PPR10, whose binding defines mRNA segments derived from two transcription units in maize chloroplasts. PPR10 interacts in vivo and in vitro with two intergenic RNA regions of similar sequence. The processed 50 and 30 RNA termini in these regions overlap by approximately 25 nucleotides. The PPR10-binding sites map precisely to these overlapping sequences, and PPR10 is required specifically for the accumulation of RNAs with these termini. These findings show that PPR10 serves as a barrier to RNA decay from either the 50 or 30 direction and that a bound protein provides an alternative to an RNA hairpin as a barrier to 30 exonucleases. The results imply that protein ‘caps’ at both 50 and 30 ends can define the termini of chloroplast mRNA segments. These results, together with recent insights into bacterial RNA decay, suggest a unifying model for the biogenesis of chloroplast transcript populations and for the determinants of chloroplast mRNA stability. The EMBO Journal (2009) 28, 2042–2052. doi:10.1038/ emboj.2009.121; Published online 7 May 2009 Subject Categories: RNA; plant biology