title: RNA-level unscrambling of fragmented genes in Diplonema mitochondria.
abstract: We previously reported a unique genome with systematically fragmented genes and gene pieces dispersed across numerous circular chromosomes, occurring in mitochondria of diplonemids. Genes are split into up to 12 short fragments (modules), which are separately transcribed and joined in a way that differs from known trans-splicing. Further, cox1 mRNA includes six non-encoded uridines indicating RNA editing. In the absence of recognizable cis-elements, we postulated that trans-splicing and RNA editing are directed by trans-acting molecules. Here, we provide insight into the post-transcriptional processes by investigating transcription, RNA processing, trans-splicing and RNA editing in cox1 and at a newly discovered site in cob. We show that module precursor transcripts are up to several thousand nt long and processed accurately at their 5' and 3' termini to yield the short coding-only regions. Processing at 5' and 3' ends occurs independently, and a processed terminus engages in trans-splicing even if the module's other terminus is yet unprocessed. Moreover, only cognate module transcripts join, though without directionality. In contrast, module transcripts requiring RNA editing only trans-splice when editing is completed. Finally, experimental and computational analyses suggest the existence of RNA trans-factors with the potential for guiding both trans-splicing and RNA editing.
authors: Department of Biochemistry, Université de Montréal, Montreal, QC, Canada.
authors: Georgette N
authors: GN
authors: Kiethega
authors: Yifei
authors: Y
authors: Yan
authors: Marcel
authors: M
authors: Turcotte
authors: Gertraud
authors: G
authors: Burger
mesh_terms: D001483:Base Sequence; D002875:Chromosomes / Q000235:genetics / Q000378:metabolism; D051545:Cyclooxygenase 1 / Q000235:genetics / Q000378:metabolism; D056898:Euglenozoa / Q000235:genetics*; D050259:Genes, Mitochondrial; D017125:Genes, Protozoan; D008928:Mitochondria / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D026723:Polyadenylation; D017393:RNA Editing; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics / Q000378:metabolism*; D020040:Trans-Splicing; D014158:Transcription, Genetic
chemical_list: D012333:RNA, Messenger; D016053:RNA, Protozoan; D051545:Cyclooxygenase 1
keywords: Diplonema papillatum; Euglenozoa; U-insertion RNA editing; trans-splicing