Journal Article

Targeted Gene Disruption Identifies Three PPR-DYW Proteins Involved in RNA Editing for Five Editing Sites of the Moss Mitochondrial Transcripts

Shotaro Ohtani, Mizuho Ichinose, Eiji Tasaki, Yoshiaki Aoki, Yoshihiro Komura and Mamoru Sugita

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 51, issue 11, pages 1942-1949
Published in print November 2010 | ISSN: 0032-0781
Published online September 2010 | e-ISSN: 1471-9053 | DOI: https://dx.doi.org/10.1093/pcp/pcq142
Targeted Gene Disruption Identifies Three PPR-DYW Proteins Involved in RNA Editing for Five Editing Sites of the Moss Mitochondrial Transcripts

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In plant organelles, RNA editing frequently occurs in many transcripts, but little is known about its molecular mechanism. Eleven RNA editing sites are present in the moss Physcomitrella patens mitochondria. Recently PpPPR_71, one member of 10 DYW-subclass pentatricopeptide repeat (PPR-DYW) proteins, has been identified as a site-specific recognition factor for RNA editing in the mitochondrial transcript. In this study, we disrupted three genes encoding a PPR-DYW protein—PpPPR_56, PpPPR_77, and PpPPR_91—to investigate whether they are involved in RNA editing. Transient expression of an N-terminal amino acid sequence fused to the green fluorescent protein (GFP) suggests that the three PPR-DYW proteins are targeted to mitochondria. Disruption of each gene by homologous recombination revealed that PpPPR_56 was involved in RNA editing at the nad3 and nad4 sites, PpPPR_77 at the cox2 and cox3 sites, and PpPPR_91 at the nad5-2 site in the mitochondrial transcripts. The nucleotide sequences surrounding the two editing sites targeted by a single PPR-DYW protein share 42 to 56% of their identities. Thus, moss PPR-DYW proteins seem to be site-specific factors for RNA editing in mitochondrial transcripts.

Keywords: DYW domain; mitochondria; Physcomitrella patens; PPR protein; RNA editing

Journal Article.  4716 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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