The force-dependent filamin A-G3BP1 interaction regulates phase-separated stress granule formation

Filamin A (FLNA) is an actin crosslinking protein that mediates mechanotransduction. External and internal mechanical forces, through the actin cytoskeleton, can induce conformational changes of the FLNA molecule to expose cryptic binding sites for its binding partners. Here, we identified G3BP1 (Ras-GTPase activating protein SH3 domain-binding protein 1) as a new FLNA mechanobinding partner. Unlike other FLNA-binding partners to the mechanosensing domain repeat 21 (R21), G3BP1 requires an additional neighboring R22 to interact. We demonstrated that their interaction occurs in the cytosol of living cells in an actin polymerization-dependent manner. We also mapped the FLNA-binding site on G3BP1 and found that a F360A point mutation in the RNA recognition motif disrupts the interaction. RNA interferes with the FLNA-G3BP1 interaction and FLNA does not localize in RNA-rich stress granules (SGs). Disruption of the interaction is sufficient to promote phase-separated SG formation and arsenite treatment further stimulates formation of SGs. Taken together, these data identify G3BP1 as a new mechanobinding protein that interacts with the FLNA mechanosensing domain R21 and suggest that SG formation is partially regulated by mechanical force.