Thiolutin Extends Replicative Lifespan by Rewiring Yeast Transcription and Metabolism

This study examines the global transcriptomic response of Saccharomyces cerevisiae strain BY4741 to short-term exposure to thiolutin, a transcriptional inhibitor. Exponentially growing cells were treated with 3 µg/mL thiolutin for 3 hours and compared with untreated controls. RNA sequencing was performed using paired-end libraries prepared with the MGIEasy RNA Library Prep Set and sequenced on the BGISEQ-500 platform. Three independent biological replicates were analyzed per condition. Differential expression analysis using DESeq2 (Benjamini–Hochberg false discovery rate < 0.05) revealed extensive transcriptional remodeling affecting pathways related to ribosome biogenesis, oxidative phosphorylation, proteostasis, stress response, and aging-associated processes. Overall design: Wild-type Saccharomyces cerevisiae BY4741 cells were cultured in YPD medium at 28°C to exponential phase. Cultures were divided into untreated controls and cells treated with 3 µg/mL thiolutin for 3 hours. Total RNA was isolated from three independent biological replicates per condition. Poly(A)-selected RNA libraries were prepared and sequenced using paired-end DNBSeq technology on the BGISEQ-500 platform. Gene-level read counts were generated and analyzed using DESeq2 to identify differentially expressed genes.