Distinct mechanical regulation of PFK1 isoforms during cytoskeletal remodeling

Abstract

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.