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

被引:28
作者
Kuznetsova, Irina [1 ,2 ]
Siira, Stefan J. [1 ,2 ]
Shearwood, Anne-Marie J. [1 ,2 ]
Ermer, Judith A. [1 ,2 ]
Filipovska, Aleksandra [1 ,2 ,3 ]
Rackham, Oliver [1 ,2 ,3 ]
机构
[1] Univ Western Australia, Harry Perkins Inst Med Res, Nedlands, WA 6009, Australia
[2] Univ Western Australia, Ctr Med Res, Nedlands, WA 6009, Australia
[3] Univ Western Australia, Sch Mol Sci, Crawley, WA 6009, Australia
基金
澳大利亚研究理事会; 英国医学研究理事会;
关键词
INTEGRATIVE GENOMICS VIEWER; MESSENGER-RNAS; POLYADENYLATION; TRANSCRIPTOME; PROTEINS; INVOLVEMENT; EXPRESSION; DECAY;
D O I
10.1093/nar/gkx104
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:5487 / 5500
页数:14
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