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Publication : Simultaneous processing and degradation of mitochondrial RNAs revealed by circularized RNA sequencing.

First Author  Kuznetsova I Year  2017
Journal  Nucleic Acids Res Volume  45
Issue  9 Pages  5487-5500
PubMed ID  28201688 Mgi Jnum  J:249591
Mgi Id  MGI:5922698 Doi  10.1093/nar/gkx104
Citation  Kuznetsova I, et al. (2017) Simultaneous processing and degradation of mitochondrial RNAs revealed by circularized RNA sequencing. Nucleic Acids Res 45(9):5487-5500
abstractText  Mammalian mitochondrial RNAs are unique as they are derived from primary transcripts that encompass almost the entire mitochondrial genome. This necessitates extensive processing to release the individual mRNAs, rRNAs and tRNAs required for gene expression. Recent studies have revealed many of the proteins required for mitochondrial RNA processing, however the rapid turnover of precursor RNAs has made it impossible to analyze their composition and the hierarchy of processing. Here, we find that circularization of RNA prior to deep sequencing enables the discovery and characterization of unprocessed RNAs. Using this approach, we identify the most stable processing intermediates and the presence of intermediate processing products that are partially degraded and polyadenylated. Analysis of libraries constructed using RNA from mice lacking the nuclease subunit of the mitochondrial RNase P reveals the identities of stalled processing intermediates, their order of cleavage, and confirms the importance of RNase P in generating mature mitochondrial RNAs. Using RNA circularization prior to library preparation should provide a generally useful approach to studying RNA processing in many different biological systems.
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