RNA editing status of nad7 intron domains in wheat mitochondria.
The most highly conserved structures of group II introns are the helical domains V and VI near the 3'splice site. Within this region of each of the four introns in the wheat mitochondrial nad7 gene encoding NADH dehydrogenase subunit 7, there are A-C mispairs. To determine whether C-to-U type RNA editing restores conventional A-U pairing, we sequenced RT-PCR products from partially-spliced nad7 template RNA and gel-fractionated, excised intron RNA. We examined transcripts from germinating wheat embryos and seedlings because these two stages of development show pronounced differences in steady state levels of nad7 intronic RNAs. We observed editing at only two of the six predicted sites, and they were located at homologous positions within domain V of the third and fourth introns. A third site was found to be edited within the unmodelled domain VI loop of the fourth intron. Similar patterns of RNA editing were seen in wheat embryos and seedlings. These observations, and the presence of other non-conventional base pairs particularly within domain V of plant mitochondrial introns, indicate weaker helical core structure than in ribozymic group II introns. Moreover, the incompleteness or absence of editing in wheat nad7 excised intron RNA suggests that, although editing may contribute to splicing efficiency, it is not essential for splicing.
Carrillo
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Biology Department, University of Ottawa, 30 Marie Curie, Ottawa K1N 6N5, Canada.
Bonen
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D001482:Base Composition; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers; D004272:DNA, Mitochondrial / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D017343:Genes, Plant / Q000235:genetics; D007438:Introns / Q000235:genetics*; D008969:Molecular Sequence Data; D009245:NADH Dehydrogenase / Q000235:genetics*; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction; D017393:RNA Editing / Q000235:genetics*; D012326:RNA Splicing / Q000235:genetics; D018749:RNA, Plant / Q000378:metabolism; D016415:Sequence Alignment; D017421:Sequence Analysis; D014908:Triticum / Q000378:metabolism*
D017931:DNA Primers; D004272:DNA, Mitochondrial; D018749:RNA, Plant; D009245:NADH Dehydrogenase