Bifurcation of stress granule fates under glycolytic inhibition enabled by association of OXPHOS gene transcripts with G3BP1

Stress granules (SGs) are dynamic organelles that form under cellular stress and are generally regarded as protective entities. However, their role in pathogenesis is increasingly recognized, yet the underlying mechanisms remain elusive due to the diversity of stress types and biological contexts. Here, we investigate how different regimes of glycolytic inhibition influence SG dynamics and transcriptomic partitioning. We subjected cells to glucose depletion (GD), 2-deoxy-D-glucose addition (2DG), or their combination (GD+2DG). We show that SGs formed under single assaults dissipate during prolonged treatment and correlate with integrated stress response (ISR), whereas SGs induced by GD+2DG persist and are associated with mitochondrial dysfunction and increased apoptosis. Using G3BP1-APEX2 proximity labeling, we find that GD+2DG uniquely enriches oxidative phosphorylation (OXPHOS) transcripts in the G3BP1-associated transcriptome. We demonstrate that mitochondrial inhibition renders SGs formed under single assaults persistent. We propose a model in which two counteracting feedback loops determine SG fates under glycolytic stress: a negative ISR-mediated loop promotes SG recovery, while a positive loop involving G3BP1-partitioned OXPHOS transcripts drives SG persistence and metabolic collapse. This framework provides mechanistic insights into the paradoxical duality of SGs in stress adaptation and disease.