Thiolutin extends replicative lifespan by rewiring yeast transcription and metabolism.

Transcription is a major cellular energy sink tightly coupled to growth, metabolism, and aging. Thiolutin, a widely used RNA polymerase inhibitor in yeast, has unclear long-term effects on aging. We show that thiolutin oppositely affects replicative and chronological aging in Saccharomyces cerevisiae by remodeling transcription, metabolism, and proteostasis(definition). Thiolutin extends replicative lifespan, increasing reproductive potential and prolonging the mitotic phase, despite slower growth and lower ATP. This longevity is linked to global transcriptional repression, including reduced ribosome biogenesis, translation, and mitochondrial oxidative phosphorylation, and dampening of TOR1-dependent growth programs. In parallel, thiolutin triggers a selective adaptive response with activation of RPN4-mediated proteasome remodeling and redox-responsive genes, without HOG1 induction. Conversely, thiolutin accelerates early chronological aging: post-mitotic survival drops alongside repression of reserve carbohydrate genes (GPH1, GSY2, TSL1), suggesting impaired adaptation when entering stationary phase. FT-Raman spectroscopy confirms coordinated depletion of RNA, proteins, lipids, and carbohydrates. Thus, thiolutin promotes a low-energy, stress-adaptive state that benefits budding yeast cells but compromises early survival of non-budding populations, underscoring transcription–energy coupling in aging trajectories.