Poster & Lightening Talk Inaugural Australian Ubiquitin Summit 2025

Molecular mechanisms of hyperosmotic stress response mediated by proteasomal degradation (128778)

Kouta Nakadai 1 , Hikaru Tsuchiya 2 , Takuya Tomita 1 , Yasushi Saeki 1
  1. The University of Tokyo, Tokyo, Japan
  2. Juntendo University, Tokyo, Japam

Acute hyperosmotic stress rapidly dehydrates cells, causing shrinkage and increasing molecular crowding, which impacts proteostasis, transcription, and translation. In response, stress-responsive MAPKs activate, modulating gene expression and translation to promote cellular adaptation. We previously reported that the ubiquitin-proteasome system (UPS) facilitates cell survival by efficiently degrading abnormal proteins generated by hyperosmotic stress through liquid-liquid phase separation. Here, we uncovered a novel hyperosmotic stress response pathway regulated by proteasomal degradation. Deep proteomic analysis revealed extensive proteome remodeling under hyperosmotic stress (approximately 150 proteins downregulated, 22 upregulated). Intriguingly, we identified a linker histone H1.2 as a substrate rapidly and specifically degraded during this stress. H1.2 is extensively ubiquitylated upon hyperosmotic stimulation, modified with complex heterotypic ubiquitin chains (K11, K29, K48 linkages). A fraction of H1.2 localized to proteasome droplets and underwent rapid, p97-dependent proteasomal degradation. We then identified the CRL4-DCAF X E3 ubiquitin ligase complex as promoting H1.2 ubiquitylation. Notably, DCAF X knockdown, which inhibited H1.2 degradation, suppressed the expression of over 200 stress-responsive genes and significantly increased cellular vulnerability to hyperosmotic stress. These findings suggest that hyperosmotic stress-dependent degradation of histone H1.2 plays a crucial role in the cellular stress response by inducing stress-responsive genes, providing a deeper understanding of how cells adapt to and survive osmotic challenges.