The cell volume regulatory response to a hypotonic stimulus is frequently initiated by activation of K+ and Cl- channels. We have characterized the hypotonic cell volume regulatory response of human melanoma cells devoid of actin-binding protein (ABP) and their genetically rescued counterpart transfected with the cDNA for ABP. ABP-deficient cells were unable to volume-regulate or activate K+ channels when exposed to a hypotonic stimulus. Genetic rescue with ABP resulted in recovery of both the cell volume regulatory response and the osmotically linked K+ channel activation. These data are consistent with a functional interaction between the actin cytoskeleton and osmotically sensitive ion transport.