The Mitochondrial S7 Ribosomal Protein Gene: Impact of DNA Rearrangements on RNA Expression in Grasses

• Main Author: Byers, Evan
• Language: en
• Published: 2012
• Subjects: Mitochondria; Ribosomal protein; RNA processing; Editing; Grasses
• Online Access: http://hdl.handle.net/10393/20516

Frequent rearrangements, typically through homologous recombination in plant mitochondrial genomes often result in different upstream and downstream sequences for the same gene among a number of species. Transcription and RNA processing signals are therefore different, even among closely related plants. To evaluate the impact of DNA rearrangements on gene expression I conducted a comparative analysis of the S7 ribosomal protein gene (rps7) among a number of grasses: wheat, rice, maize, barley, rye, brome, Lolium and oats (grasses whose evolutionary divergence times range from about 5 to 60 Mya). Using circularized-RT-PCR to simultaneously map rps7 transcript termini I found that 3’ends for various RNA species are homogeneous, mapping to conserved sequences among plants. 5’ termini are more complex and can be both discrete and heterogeneous for different transcripts, both within and among plants. Genome rearrangements upstream of the rps7 start codon for some but not all species has led to plant-specific signals for both rps7 transcription and RNA processing. Termini for rps7 precursor species in wheat and Lolium are very discrete and likely use different upstream tRNAs as processing signals for end-cleavage. A number of potential stem-loop structures have also been identified at or near 5’ and 3’ termini which may function in maturation of transcript ends or provide transcript stability and protection from degradation by ribonucleases. C-to-U RNA editing of non-coding sequences, a rare event, was observed at multiple sites within the 5’ and 3’UTRs among plants. Some sites may even be developmentally regulated as CR-RT-PCR experiments were conducted using mitochondrial RNA isolated from seedlings and germinating embryos. Taken together, my observations demonstrate the frequency of upstream DNA rearrangements and the variety of signals used for expression of rps7 among grasses, providing new insights into the complexities of mRNA production in plant mitochondria.