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Analysis of mitochondrial RNA-processing defects in patient-derived tissues by qRT-PCR and RNAseq.
Methods Mol. Biol. 1567, 379-390 (2017)
Transcription of the mitochondrial genome yields three large polycistronic transcripts that undergo multiple endonucleolytic processing steps, before resulting in functional mRNAs, tRNAs, and rRNAs. Cleavage of the large precursor transcripts is mainly performed by the RNase P complex and RNase Z that cleave mitochondrial pre-tRNAs at their 5′ and 3′ ends respectively. Most likely there are additional enzymes involved that still await identification and characterization. Defects in mitochondrial RNA processing have been associated with human disease. There are published cases of patients carrying mutations in either HSD17B10/MRPP2 (encoding a subunit of RNase P complex) or ELAC2 (coding for RNase Z). In addition, several mtDNA mutations within tRNA genes have been shown to affect RNA processing. Here, we describe detailed protocols for analyzing RNA processing of mitochondrial tRNAs, in particular their 3′-ends that are processed by RNase Z. These protocols should serve as a guide to extract RNA for quantitative real-time PCR and RNAseq analysis.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Elac2 ; Mitochondrial Rna Processing ; Mrpp2 ; Mtdna ; Qrt-pcr ; Rnase P ; Rnase Z ; Rnaseq
ISSN (print) / ISBN 1064-3745
Conference Title Mitochondria
Journal Methods in Molecular Biology
Quellenangaben Volume: 1567, Pages: 379-390
Publishing Place Berlin [u.a.]
Reviewing status Peer reviewed
Institute(s) Institute of Human Genetics (IHG)