Long-read direct RNA sequencing of the mitochondrial transcriptome of Saccharomyces cerevisiae

Mitochondria fulfil many essential roles in eukaryotic cells, yet some of their molecular mechanisms are still unexplored. Although 99% of the mitochondrial proteins are imported from the cytosol, mitochondria have their own DNA, transcription and translation machinery. The Saccharomyces cerevisiae mitochondrial DNA contains 11 polycistronic transcripts that encode 2 ribosomal subunits, 24 tRNAs and 9 genes, which can be spliced in alternative ways to yield different proteins. There are still many unresolved questions about mitochondrial genes and their splicing, including how gene expression and splicing is affected by different growth conditions and what role introns play in mitochondrial physiology. In the present study, we aimed to elucidate this by developing an RNA-sequencing method for mitochondrial RNA using Nanopore technology. Overall design: Comparision of transcription between the mitochondrial transcriptomes of cells grown on glucose versus cells grown on ethanol. All data is derived from mitchondria isolated from biological triplicate cultures and sequenced with Nanopore Direct RNA sequencing