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Large-scale detection and analysis of RNA editing in grape mtDNA by RNA deep-sequencing

Author:
Picardi, Ernesto, Horner, David S., Chiara, Matteo, Schiavon, Riccardo, Valle, Giorgio, Pesole, Graziano
Source:
Nucleic acids research 2010 v.38 no.14 pp. 4755-4767
ISSN:
0305-1048
Subject:
RNA editing, Vitis vinifera, animals, genes, grapes, high-throughput nucleotide sequencing, mitochondria, mitochondrial DNA, non-coding RNA, plants (botany), proteomics
Abstract:
RNA editing is a widespread post-transcriptional molecular phenomenon that can increase proteomic diversity, by modifying the sequence of completely or partially non-functional primary transcripts, through a variety of mechanistically and evolutionarily unrelated pathways. Editing by base substitution has been investigated in both animals and plants. However, conventional strategies based on directed Sanger sequencing are time-consuming and effectively preclude genome wide identification of RNA editing and assessment of partial and tissue-specific editing sites. In contrast, the high-throughput RNA-Seq approach allows the generation of a comprehensive landscape of RNA editing at the genome level. Short reads from Solexa/Illumina GA and ABI SOLiD platforms have been used to investigate the editing pattern in mitochondria of Vitis vinifera providing significant support for 401 C-to-U conversions in coding regions and an additional 44 modifications in non-coding RNAs. Moreover, 76% of all C-to-U conversions in coding genes represent partial RNA editing events and 28% of them were shown to be significantly tissue specific. Solexa/Illumina and SOLiD platforms showed different characteristics with respect to the specific issue of large-scale editing analysis, and the combined approach presented here reduces the false positive rate of discovery of editing events.
Agid:
2272326