Distinct mechanical regulation of PFK1 isoforms during cytoskeletal remodeling

All cells experience mechanical forces that shape their behavior and organization. These forces are sensed at cell-cell adhesions and transduced through the actin cytoskeleton, triggering cytoskeletal reinforcement to withstand mechanical load. Such remodeling is energy-intensive and supported by increased glycolysis, yet the mechanisms linking mechanical stress to metabolic activation remain incomplete. Here, we identify phosphofructokinase-1 (PFK1) as a bridge coupling mechanotransduction and metabolism. Shear stress activates PFK1 and promotes its association with F-actin, enhancing reinforcement, glucose uptake, and intracellular ATP generation. Among the three human PFK1 isoforms, only the muscle isoform (PFKM) is essential for this response. The PFKM C-terminal tail is required for complete actin engagement and mechanosensitive activation: deletion abolishes this function, while grafting the tail onto other isoforms restores it. These findings establish PFKM as a mechanosensitive metabolic enzyme that links cytoskeletal mechanics to energy production, providing a framework for how mechanical and metabolic dysfunctions intersect in disease.