Identification of critical nucleotide positions for plastid RNA editing site recognition

Transcripts in higher plant cell organelles undergo RNA editing by C-to-U conversion. Both the mechanistic steps and the factors involved in this process are largely unknown. To gain a better understanding of the molecular interactions involved in organellar RNA editing, we have begun to identify cr...

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Bibliographic Details
Published in:RNA (Cambridge) 1997-10, Vol.3 (10), p.1194-1200
Main Authors: Bock, R, Hermann, M, Fuchs, M
Format: Article
Language:English
Subjects:
Biolistics
Chloroplasts - genetics
Chloroplasts - metabolism
Cytosine - metabolism
Mutagenesis
Nicotiana - genetics
Nucleotides - chemistry
Nucleotides - genetics
Nucleotides - metabolism
Oligodeoxyribonucleotides - chemistry
Plant Proteins - genetics
Plants, Toxic
Point Mutation
Polymerase Chain Reaction
RNA Editing
RNA, Messenger - chemistry
RNA, Messenger - genetics
RNA, Messenger - metabolism
RNA, Plant - chemistry
RNA, Plant - genetics
RNA, Plant - metabolism
Sequence Deletion - genetics
Transcription, Genetic
Transformation, Genetic
Uracil - metabolism
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Summary:Transcripts in higher plant cell organelles undergo RNA editing by C-to-U conversion. Both the mechanistic steps and the factors involved in this process are largely unknown. To gain a better understanding of the molecular interactions involved in organellar RNA editing, we have begun to identify critical nucleotide positions for plastid RNA editing-site recognition. We performed a scanning point mutagenesis on a sequence motif separating editing sites IV and V in the tobacco ndhB transcript. The constructs were integrated into the chloroplast genome by the biolistic process and the effect of each point mutation on editing of both the upstream and the downstream site was measured. In addition to a previously identified sequence element located upstream of both sites, only few nucleotide positions 5' and 3' of an editing site turned out to be of critical importance. Unexpectedly, our study revealed that mutation of the upstream site leads to loss of editing at the downstream site. However, our results also indicate that, even though closely adjacent editing sites can share common recognition elements in cis, they are edited independently and not in a polar fashion.
ISSN:1355-8382
1469-9001