Impact of genomic environment on mitochondrial rps7 mRNA features in grasses

Abstract

The mitochondrial genomes of flowering plants are highly recombinogenic and this can lead to altered transcriptional units, even between closely related species. We are interested in the effects that DNA rearrangements have on the generation of mature mRNAs, and to this end we have determined the termini of mitochondrial S7 ribosomal protein (rps7) mRNAs from selected grasses, using circularized-RT-PCR. Although the rps7 mRNAs show a similar size of about 750 nt by northern hybridization analysis and have virtually identical 3' UTRs, their 5' terminal extremities differ among plant species, and this is attributable to genome rearrangements in some but not all cases. In wheat, rice, and barley, the 5' ends are homogeneous for each plant but map to non-homologous sites among the three species. In contrast, the rye, brome and Lolium 5' ends are quite heterogeneous in length even though they are located within conserved genomic regions. Comparative sequence analysis suggests that certain grass lineages have retained an ancestral organization upstream of rps7 that includes a 170-bp block homologous to sequences preceding several other mitochondrial genes, whereas others have undergone independent rearrangements at a recombination-prone site. Our analysis of mature rps7 transcripts revealed two non-silent RNA edits within the coding sequences, and also editing at several sites within the conserved 5' and 3' UTR regions in these plants, raising the possibility of their role in rps7 expression at the post-transcriptional level. Taken together, our observations illustrate the dynamic nature of upstream regulatory cis-elements for mitochondrial rps7 mRNA production in contrast to conservative 3' end-formation signals, during evolution in grasses.