Phosphorylation of human fascin inhibits its actin binding and bundling activities.

Human fascin is an actin-bundling protein that is thought to be involved in the assembly of actin filament bundles present in microspikes as well as in membrane ruffles and stress fibers. We have found that human fascin is phosphorylated in vivo upon treatment with 12-O-tetradecanoylphorbol-13-acetate, a tumor promoter. The in vivo phosphorylation is gradually increased from 0.13 to 0.30 mol/mol during 2 h of treatment, concomitant with disappearance of human fascin from stress fibers, membrane ruffles, and microspikes. Human fascin can also be phosphorylated in vitro as judged by phosphopeptide mapping. The extent of phosphorylation depends on pH: the stoichiometries are 0.05, 0.38, and 0.6 alone does not affect fascin-actin binding. With the incorporation of 0.25 mol of phosphate/mol of protein, the actin binding affinity is reduced from 6.7 x 10(6) to 1.5 x 10(6) m(-1). The actin bundling activity is also decreased. These results suggest that phosphorylation of fascin plays a role in actin reorganization after treatment with 12-O-tetradecanoylphorbol-13-acetate.