Assays for investigating RNA editing in plant mitochondria.
In plant organelles transcripts are modified posttranscriptionally by RNA editing. This modification process changes almost every protein-coding RNA at specific cytidine and uridine positions. Therefore, mitochondrially encoded protein sequences differ from the genomically fixed information and show, after editing, a higher conservation. To investigate this unusual processing step in plant mitochondria, several assays have been developed. However, compared with the progress made in other RNA editing fields, knowledge about the factors involved in plant mitochondrial editing is limited. One reason for this is the lack of a reliable in vitro system for mitochondria. To reveal the biochemical nature of the RNA editing reaction in plant mitochondria, we developed an in vitro system by which we were able to show that cytidine is specifically modified to uridine by a deamination or transamination process. Here we describe the development of a pea in vitro system and discuss assays to follow the editing process.
Yu
W
W
Institute for Applied Genetics, Free University of Berlin, Germany.
Sánchez
H
H
Schuster
W
W
D005821:Genetic Techniques; D008928:Mitochondria / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010944:Plants / Q000235:genetics* / Q000648:ultrastructure; D012313:RNA / Q000235:genetics / Q000378:metabolism; D017393:RNA Editing; D000077278:RNA, Mitochondrial; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D013698:Templates, Genetic; D014158:Transcription, Genetic
D000077278:RNA, Mitochondrial; D018749:RNA, Plant; D012313:RNA