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.线粒体在真核细胞中发挥诸多关键功能，但其部分分子机制仍未被完全阐明。尽管99%的线粒体蛋白质均由胞质溶胶输入，但线粒体自身拥有独立的DNA、转录及翻译系统。酿酒酵母（Saccharomyces cerevisiae）的线粒体DNA包含11个多顺反子转录本，其编码2个核糖体亚基、24种转运RNA（tRNA）以及9个基因，该类转录本可通过可变剪接产生不同的蛋白质。目前关于线粒体基因及其剪接过程仍存在诸多未解决的问题，例如不同生长条件如何影响基因表达与剪接，以及内含子在线粒体生理过程中发挥何种作用。本研究旨在通过开发基于纳米孔（Nanopore）技术的线粒体RNA测序方法，对上述问题进行阐释。