Poster Presentation 44th Lorne Genome Conference 2023

Aberrant Non-Coding RNA Polyadenylation is an Early Stress Response in Saccharomyces cerevisiae (#241)

Calvin A Kraupner-Taylor 1 2 , Traude H Beilharz 1 2
  1. Development and Stem Cell Program, Monash Biomedicine Discovery Institute, Monash University, Melbourne, VIC, Australia
  2. Biochemistry and Molecular Biology, Monash University, Mount Waverley, VIC, Australia

The metabolism of non-coding RNA is becoming an increasingly complex field of research. tRNA are subject to diverse epigenetic modifications, their abundance can be adapted to the translational needs of the cell and additional regulatory roles are attributed to fragments generated during decay. These processes hold capacity to significantly diversify tRNA function from the traditional role as stable translational machinery components. Our research probes the idea that derailment of normal co-transcriptional RNA processing is a universal response to stress in S. cerevisiae. It is known that splicing and alternative polyadenylation are regulated during stress. Here, we describe a new aspect of non-coding RNA regulation which rapidly responds to environmental stress. We used an in-house whole transcriptome RNA-seq method to capture early changes to gene expression and poly(A) tail length distribution during cellular adaptation to osmotic stress. In addition to changes in mRNA expression and polyadenylation-state, we identified rapid polyadenylation of snoRNA rRNA and pre-tRNAs within 5 minutes. Unprocessed, short poly(A) tailed, pre-tRNA transcripts accumulated transiently, being removed from the transcriptome following adenylation by 15 minutes of stress. The non-canonical poly(A) polymerase Trf4p is required for this polyadenylation, suggesting tRNA adenylation is carried out by the Nuclear TRAMP complex. Interestingly, the process appears to primarily target proline and glutamine accepting tRNAs, suggesting a selective mechanism. Proline and glutamine were previously described as osmoprotectants in bacteria, and mRNA encoding the proline transporter PUT4 is upregulated by 30 minutes here. We propose the pre-tRNA polyadenylation, and metabolic uncoupling, of processing of these tRNA toward decay represents a previously uncharacterised arm of cell biology that allows yeast to rapidly adjust their intracellular environment to adapt to osmotic stress preceding adaptation through the proteome.