GSE173837 RNA-seq and small RNA-seq in fission yeast CPF mutants that enable small-RNA-mediated epigenetic gene silencing

In this study, we have combined chemical mutagenesis with whole-genome sequencing in a sensitized S. pombe reporter strain to obtain a more comprehensive list of putative suppressors of small-RNA-mediated epigenetic gene silencing. This revealed more than 20 novel silencing-enabling mutations in genes that are associated with RNA processing, regulation of transcription, or post-translational protein modification. Focusing on factors involved in pre-mRNA cleavage and polyadenylation, we for example show that single amino acid substitutions in Yth1, which is responsible for polyadenylation signal recognition, lead to nearly 100% effective de novo formation of silent heterochromatin. In S. pombe, small RNA-mediated silencing relies on the continuous amplification of the siRNA pool through a positive feedback loop. The deposited data shows the generation of secondary small RNAs over the reporter gene exclusively in the presence of RNAi-enabling mutations in the pre-mRNA cleavage and polyadenylation machinery. Furthermore, the RNA-seq data show that overall transcriptome levels are not markedly changing in the mutations themselves. Altogether, our work shows that epigenetic gene silencing can be enabled by the acquisition of a plethora of mutant alleles in fission yeast.
Source: GEO: Gene Expression Omnibus - Category: Genetics & Stem Cells Tags: Expression profiling by high throughput sequencing Non-coding RNA profiling by high throughput sequencing Schizosaccharomyces pombe Source Type: research