An in vitro RNA editing system from cauliflower mitochondria: editing site recognition parameters can vary in different plant species.
Most of the 400 RNA editing sites in flowering plant mitochondria are found in mRNAs. Consequently, the sequence vicinities of homologous sites are highly conserved between different species and are presumably recognized by likewise conserved trans-factors. To investigate the evolutionary adaptation to sequence variation, we have now analyzed the recognition elements of an editing site with divergent upstream sequences in the two species pea and cauliflower. This variation is tolerated at the site selected, because the upstream cis-elements reach into the 5'-UTR of the mRNA. To compare cis-recognition features in pea and cauliflower mitochondria, we developed a new in vitro RNA editing system for cauliflower. In vitro editing assays with deleted and mutated template RNAs show that the major recognition elements for both species are located within the conserved sequence. In cauliflower, however, the essential upstream nucleotides extend further upstream than they do in pea. In-depth analysis of single-nucleotide mutations reveals critical spacing of the editing site and the specific recognition elements, and shows that the +1 nucleotide identity is important in cauliflower, but not in pea.
Neuwirt
Julia
J
Molekulare Botanik, Universität Ulm, 89069 Ulm, Germany.
Takenaka
Mizuki
M
van der Merwe
Johannes A
JA
Brennicke
Axel
A
D020121:5' Untranslated Regions; D029681:Arabidopsis Proteins; D001483:Base Sequence; D001937:Brassica / Q000166:cytology / Q000235:genetics*; D002457:Cell Extracts; D002474:Cell-Free System / Q000737:chemistry / Q000378:metabolism; D017124:Conserved Sequence; D019143:Evolution, Molecular; D014644:Genetic Variation; D066298:In Vitro Techniques; D025261:Mitochondrial Proton-Translocating ATPases; D008969:Molecular Sequence Data; D018532:Pisum sativum / Q000166:cytology / Q000235:genetics; D017354:Point Mutation; D012313:RNA / Q000737:chemistry* / Q000378:metabolism; D017393:RNA Editing; D000077278:RNA, Mitochondrial; D018749:RNA, Plant / Q000737:chemistry* / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017384:Sequence Deletion; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D013698:Templates, Genetic
D020121:5' Untranslated Regions; C063668:ATP9 protein, Arabidopsis; D029681:Arabidopsis Proteins; D002457:Cell Extracts; D000077278:RNA, Mitochondrial; D018749:RNA, Plant; D012313:RNA; D025261:Mitochondrial Proton-Translocating ATPases